JP2005325055A - Ligand binding protein-containing complex and screening method for substance binding to ligand binding protein - Google Patents

Abstract

It is possible to avoid the complexity and low reproducibility caused by using a cell itself as a screening system, and to target only a target ligand binding protein without being influenced by other ligand binding proteins. A screening method that can be used and a novel complex that can be used for the screening method are provided.
The complex is selected from the group consisting of (1) a protein capable of specifically binding to a ligand, a fragment of the protein containing a ligand binding site, and a polypeptide-containing substance containing the protein or the fragment. A ligand-binding protein or a derivative thereof; and (2) a partner substance capable of specifically binding to the ligand-binding protein or a derivative thereof at a site other than the ligand-binding site.
[Selection figure] None

Description

  The present invention relates to a complex containing a protein that can specifically bind to a ligand (hereinafter referred to as “ligand binding protein”), and a substance that binds to the ligand binding protein (hereinafter referred to as “target substance”). The present invention relates to a screening method and a kit. The screening method and kit can also be used for analysis of binding and / or action of a target substance to a ligand-binding protein.

  A cell must quickly and accurately grasp changes in its surrounding environment and adjust the biochemical reaction and gene expression in the cell to respond to it. As an initial step, it is a ligand binding protein such as a receptor or a cell adhesion molecule present in the cell membrane that plays a role of receiving an extracellular stimulus and transmitting it to the inside of the cell. Among the receptors, one that works in combination with G protein is called a G protein-coupled receptor (hereinafter referred to as “GPCR”).

  Substances that transmit stimuli (information) to cells such as hormones and cytokines are substances that are inherent in the body and transmit information to cells. These are called (endogenous) ligands. Each ligand plays a role by binding to a specific ligand-binding protein. There are various kinds of ligands such as proteins, peptides, lipids, and light, and since the places where the ligand binding proteins are expressed are diverse, it is not possible to describe the function of the ligand binding proteins in a general way. For example, it is known to be involved in the five senses such as the immune system, cell proliferation, or auditory sense.

  In recent years, with the completion of decoding of human DNA, gene prediction and accompanying protein expression have been actively performed. Since GPCR has a characteristic structure of penetrating the cell membrane seven times, its presence can be easily predicted from DNA information. As a result, it is also suggested that there are about 200 kinds of unknown GPCRs. Such a GPCR can roughly predict the expression site by RT-PCR or the like, and can predict what type of ligand from homology.

  GPCRs are known to be involved in various physiological phenomena in vivo, and about 50% of currently marketed drugs are drugs targeting GPCRs. Therefore, GPCR may be very useful as a drug target, and many major pharmaceutical companies and venture companies around the world are conducting research and development targeting GPCR. However, there is no known method for efficiently finding agonists and antagonists as drug candidate substances, and enormous costs and time have been spent for the discovery.

When a ligand binds to the GPCR, the G protein undergoes a conformational change from the GDP-bound type to the GTP-bound type in the cell membrane. G protein is a trimeric protein composed of α, β, and γ subunits, and GTP and GDP bind to G protein α subunit. The G protein α subunit to which GTP is bound dissociates from the GPCR and the G protein β · γ subunit and acts on the effector. G protein α subunits can be broadly classified into Gs, Gi, Gq, and G _12/13 families, each having a different effector. For example, Gs activates adenylate cyclase (hereinafter referred to as AC), and Gi inactivates AC. Gq acts on phospholipase (hereinafter referred to as PLC). AC produces cAMP, a second messenger, and PLC produces inositol triphosphate (IP ₃ ) and diacylglycerol (DG).

The GPCR cascade reaction as described above further causes the behavior of cells through release of Ca ²⁺ and phosphorylation of the target protein. Many conventional screenings for ligands and agonists / antagonists utilize such a cascaded process. Since such a conventional screening technique is performed in a cell system, there are many problems such as the complexity of cell culture, the possibility of detecting reactions other than the target GPCR of the test substance, and variations in measurement due to contaminants. is there.

For example, in the case of a GPCR assay system, the following methods are known as conventional assay methods. That is, in the case of a second messenger produced by the action of an effector system, for example, a Gs or Gi-coupled receptor, by causing a test substance (test sample) to act on cells expressing the target GPCR alone or in combination with a known agonist In general, a technique for measuring the amount of cAMP linked to fluctuations in the activity of adenylate cyclase and the concentration of IP ₃ or Ca ²⁺ linked to fluctuations in the activity of phospholipase C in the case of Gq-coupled receptors are widely used. In addition, an assay method using cells expressed in the form of a fusion polypeptide of GPCR and G protein α subunit instead of expressing GPCR as it is is also known (Patent Document 1).

  However, these methods using cells require extremely complicated operations, and it has been extremely difficult to rapidly evaluate a large amount of test substances. In addition, in these methods, it is necessary to select a second messenger to be measured depending on the type of G protein to which the target GPCR is coupled. Therefore, when the G protein conjugated with the target GPCR is unknown, it is necessary to perform the assay in all assay systems capable of measuring messages via all of Gs, Gi, and Gq. Furthermore, for receptors that couple to Gi, the secondary message induced by conjugation of the target GPCR to Gi is a decrease in cAMP due to suppression of adenylate cyclase. Therefore, in order to quantitatively capture the secondary message induced by conjugation with Gi, after increasing the amount of cAMP by increasing the amount of cAMP with a reagent (forskolin or the like) that activates Gs in advance and increases cAMP in advance. Therefore, it was necessary to evaluate the amount of decrease in the amount of cAMP caused by the test substance, and a very complicated operation was required.

  As a method for avoiding the complexity and low reproducibility caused by using cells themselves as a screening system, an agonist or antagonist screening method using a cell membrane fraction containing GPCR has been developed (Non-patent Document 1, Patent). Reference 2). For example, Non-Patent Document 1 discloses a method of separating a cell membrane fraction from an animal cell in which a large amount of a target GPCR is expressed, and using the cell membrane fraction to obtain a substance that binds to the cell membrane fraction. Yes. Further, in Patent Document 2, instead of expressing a GPCR as it is, a screening method using a cell membrane system that expresses a GPCR and a G protein α subunit in the form of a fusion polypeptide and uses the fusion polypeptide. Is disclosed. However, in any method, it is clear that in addition to the target GPCR, a huge variety of receptors including known GPCRs and unknown GPCRs are contained in the cell membrane fraction, and the target GPCR is expressed in large quantities. Although attempts have been made to suppress the influence of receptors other than the target GPCR, substances that bind to receptors other than the target are often obtained, and even if candidate compounds are obtained, agonists for the target receptor Or further analysis of whether or not it is an antagonist was necessary.

  On the other hand, as a method for identifying a compound that binds to a GPCR, a partially purified GPCR is bound to a support for column chromatography via a tag linked to its end, and a peptide library is attached to a column packed with the support. Patent Document 3 discloses a method for obtaining a GPCR-binding compound by passing it through. However, although Patent Document 3 describes that a column packed with a GPCR binding support was actually prepared and passed through a peptide library, there was no specific description of the obtained GPCR binding compound. There is no support in Patent Document 3 that GPCR-binding compounds can actually be obtained by this method. Also, this method makes it impossible to determine an agonist or antagonist for the target GPCR, and further analysis can be performed using another assay system, that is, an assay system using the cell itself or a cell membrane fraction. It is necessary to carry out the above, and it does not eliminate the various disadvantages in the aforementioned assay system.

JP 2003-250551 A JP 2003-210192 A JP-T-2004-502134 “Life Sciences”, (UK), 2003, 74, 4, p. 489-508

  Therefore, the object of the present invention is to eliminate these disadvantages of the prior art, to avoid the complexity and low reproducibility caused by using the cells themselves as a screening system, and in addition to other ligand binding proteins. It is an object of the present invention to provide a screening method that can target only a target ligand-binding protein without being affected.

  Since the complex of the present invention contains an isolated ligand-binding protein, it is possible to solve the problem of the conventional technique that a substance that binds to a receptor other than the target is removed. That is, the complex of the present invention expresses a target ligand-binding protein in a large amount in, for example, insect cells, solubilizes only the ligand-binding protein from a membrane fraction obtained by disrupting the cells with a homogenizer, and then binds the ligand binding protein. It is obtained by isolating the target ligand-binding protein by utilizing the binding with a partner substance that binds to the protein, and preferably immobilizing it on a carrier. Therefore, the screening method using the complex of the present invention can accurately perform screening using only the target ligand-binding protein as a target under the condition that no other receptor exists. In addition, according to a preferred embodiment of the present invention, GPCR can be expressed in a form fused with G protein, so that it can be used for screening methods and also for measuring the activity of ligands or agonists / antagonists.

The present invention
[1] (1) (i) a protein capable of specifically binding to a ligand, (ii) a fragment of the protein containing a ligand binding site, and (iii) the protein or a polypeptide-containing substance containing the fragment A ligand binding protein or derivative thereof selected from
(2) a complex comprising a partner substance capable of specifically binding to the ligand-binding protein or a derivative thereof at a site other than the ligand-binding site;
[2] The ligand binding protein or derivative thereof is a transmembrane receptor, a fragment of the receptor containing a ligand binding site, or a polypeptide-containing substance containing the receptor or the fragment. Complex of;
[3] The ligand binding protein or derivative thereof is a nuclear receptor, a fragment of the receptor containing a ligand binding site, or a polypeptide-containing substance containing the receptor or the fragment. Complex;
[4] The ligand binding protein or derivative thereof is a G protein-coupled receptor, a fragment of the receptor containing a ligand binding site, or a polypeptide-containing substance containing the receptor or the fragment. The described complex;
[5] The complex according to [1], wherein the ligand-binding protein or a derivative thereof is hGPCR44, a fragment of the receptor including a ligand binding site, or a polypeptide-containing substance including the receptor or the fragment;
[6] The complex according to [1], wherein the ligand-binding protein or derivative thereof is human hGPCR44, a fragment of the receptor containing a ligand-binding site, or a polypeptide-containing substance containing the receptor or the fragment. ;
[7] The complex according to [1], wherein the ligand-binding protein or a derivative thereof is a fusion polypeptide of a G protein-coupled receptor or a fragment thereof and a G protein or an α subunit or a fragment thereof;
[8] The complex according to [1], wherein the ligand-binding protein or derivative thereof is a fusion polypeptide of hGPCR44 or a fragment thereof and G protein or an α subunit or a fragment thereof;
[9] The complex according to [1], wherein the ligand-binding protein or derivative thereof is a fusion polypeptide of human hGPCR44 or a fragment thereof and G protein or an α subunit or a fragment thereof;
[10] The complex according to any one of [1] to [9], wherein the partner substance is an antibody, biotin, or avidin;
[11] The complex according to any one of [1] to [9], wherein the partner substance is an antibody;
[12] The complex according to any one of [1] to [9], wherein the partner substance is an IgG antibody;
[13] The complex according to any one of [1] to [9], wherein the partner substance is a Fab fragment of an IgG antibody;
[14] The complex according to any one of [1] to [9], wherein the partner substance is a fragment containing an antigen recognition site of an IgG antibody;
[15] A screening method for a substance that binds to a ligand-binding protein, which comprises contacting the complex according to any one of [1] to [14] with a test substance;
[16] (i) a step of bringing the complex according to any one of [1] to [14] supported on a carrier into contact with a test substance, and (ii) the above-described complex via the complex The screening method according to [15], comprising a step of obtaining a test substance bound to a carrier;
[17] (i) a step of contacting the test substance with the complex according to any one of [1] to [14] supported on a carrier via a partner substance, and (ii) the complex The screening method according to [16], comprising a step of obtaining a test substance bound to the carrier via a body;
[18] (1) (1a) Ligand binding protein selected from the group consisting of a protein capable of specifically binding to a ligand, a fragment of the protein containing a ligand binding site, and a polypeptide-containing substance containing the protein or the fragment Or a derivative thereof, (1b) a polynucleotide comprising a base sequence encoding the amino acid sequence of the ligand binding protein or derivative thereof, or (1c) a vector comprising the polynucleotide,
(2) a kit for screening a substance that binds to the ligand-binding protein, comprising a partner substance that can specifically bind to the ligand-binding protein or a derivative thereof at a site other than the ligand-binding site;
[19] A kit for screening a substance that binds to the ligand-binding protein, comprising the complex according to any one of [1] to [14];
[20] Binding and / or action of the test substance to the ligand binding protein, wherein the test substance is brought into contact with the complex according to any one of [1] to [14]. How to analyze;
[21] (i) A step of bringing the complex according to any one of [7] to [9] supported on a carrier into contact with a test substance in the presence of labeled GTP, and (ii) ) A method for analyzing the binding and / or action of the test substance to a receptor, comprising the step of analyzing the binding of GTP to the complex;
[22] (i) A step of bringing the complex according to any one of [7] to [9] supported on the plate into contact with a test substance in the presence of labeled GTP, and (ii) ) A method for analyzing the binding and / or action of the test substance to a receptor, comprising the step of analyzing the binding of GTP to the complex;
[23] (i) contacting the complex according to any one of [7] to [9] supported on a plate with a peptide-displayed phage in the presence of labeled GTP; and (ii) ) A method for analyzing the binding and / or action of the peptide-displayed phage to a receptor comprising the step of analyzing the binding of GTP to the complex;
[24] (1) (1a) Ligand binding protein selected from the group consisting of a protein capable of specifically binding to a ligand, a fragment of the protein containing a ligand binding site, and a polypeptide-containing substance containing the protein or the fragment Or a derivative thereof, (1b) a polynucleotide comprising a base sequence encoding the amino acid sequence of the ligand binding protein or derivative thereof, or (1c) a vector comprising the polynucleotide,
(2) A binding and / or action of a test substance to the ligand-binding protein, comprising a partner substance capable of specifically binding to the ligand-binding protein or a derivative thereof at a site other than the ligand-binding site. Analysis kit;
[25] A kit for analysis of binding and / or action of a test substance to a ligand-binding protein, comprising the complex according to any one of [1] to [14].

  According to the complex or screening method of the present invention, the complexity and low reproducibility caused by using the cell itself as a screening system can be avoided, and without being affected by other ligand-binding proteins, Only the target ligand-binding protein can be targeted, and screening can be performed with a high S / N ratio (for example, the ratio of signals detected before and after stimulation with a test substance).

Further, according to a preferred embodiment of the present invention using GPCR as a ligand binding protein, GPCR or a fragment thereof (particularly GPCR) is fused with G protein or an α subunit thereof or a fragment thereof (particularly G protein or an α subunit thereof). By expressing in a form, it can be used for measuring the activity of a ligand or agonist / antagonist obtained by screening.
Furthermore, the complex of the present invention contains a carrier, and further, as a fusion polypeptide of GPCR or a fragment thereof and G protein or an α subunit or a fragment thereof, the C-terminus of GPCR or a fragment thereof (particularly GPCR), When using a fusion polypeptide in which the N-terminus of a protein or its α subunit or a fragment thereof (particularly G protein or its α subunit) is used, the ligand binding site can be located away from the carrier. The contact opportunity between the test substance and the ligand binding site can be increased. For example, when the carrier is a particle, the ligand binding site is arranged toward the outside of the particle.

  In addition, since the complex of the present invention can be screened using the binding between the complex of the present invention and the test substance, the test substance that does not bind to the test substance that binds to the complex of the present invention Can be separated using a carrier carrying the complex of the present invention, so that a mixture of a plurality of different test substances can be directly used for screening. For example, when the test substance is a phage library of a peptide-displayed phage, the peptide binding to the ligand-binding protein can be screened by contacting the phage library with one ligand-binding protein. Therefore, the screening method using the complex of the present invention does not need to carry out a huge number of screenings for each test substance, which is necessary in the conventional method, and enables efficient screening.

  Furthermore, as described above, the complex of the present invention can separate a test substance that binds and a test substance that does not bind to the target ligand binding protein using a carrier or the like. It can also be used for purification.

[1] Complex and kit of the present invention
(1) selected from the group consisting of (i) a protein capable of specifically binding to a ligand, (ii) a fragment of the protein containing a ligand binding site, and (iii) a polypeptide-containing substance containing the protein or the fragment A ligand binding protein or derivative thereof;
(2) A complex comprising a partner substance capable of specifically binding to the ligand-binding protein or derivative thereof.

“Ligand-binding protein or derivative thereof”, which is the first component forming the complex of the present invention,
(I) a ligand binding protein,
It can be selected from the group consisting of (ii) a fragment of the ligand binding protein containing a ligand binding site, and (iii) a polypeptide-containing substance containing the ligand binding protein (i) or the fragment (ii). Hereinafter, unless otherwise specified, the term “fragment of a ligand binding protein” means a fragment of a ligand binding protein and containing a ligand binding site, unless otherwise specified.

The “ligand binding protein” is not particularly limited as long as it is a protein that binds to a ligand and exhibits an action in vivo, and examples thereof include a receptor or a cell adhesion molecule.
Examples of the receptor include a receptor present on the cell surface (transmembrane receptor) or a receptor present in the cell (for example, a nuclear receptor). The receptor is a naturally occurring receptor [preferably a mammal (eg, human, mouse, rat, dog, cat, monkey, pig, cow, sheep, goat, horse, or dolphin, particularly human). Derived receptor] or a modified receptor thereof.

  Examples of the transmembrane receptor include an ion channel-linked receptor or a receptor that is not linked to an ion channel, and examples thereof include a kinase type receptor or a non-kinase type receptor. Can do.

The ion channel-coupled receptor is a small pore receptor penetrating the cell membrane, and the small pore opens and closes by the binding of a ligand molecule as a signal, and Na ⁺ , K ⁺ , Alternatively, ion transport such as Ca ²⁺ is performed. Examples of the ion channel-linked receptor include each receptor having acetylcholine, glutamic acid, serotonin, GABAa, or glycine as a ligand. For example, an ion channel receptor having acetylcholine, glutamic acid, or serotonin as a ligand transports Na ⁺ , and an ion channel receptor having GABAa or glycine as a ligand transports Cl ⁻ .

  The kinase type receptor is a receptor having a single-penetration structure and having a kinase bound to the cell membrane. Examples of the kinase type receptor include a growth factor type receptor family or a TGF (transforming growth factor) -β receptor family.

  The growth factor receptor family has tyrosine kinases inside the cell membrane, and is a receptor that is activated by phosphorylation of tyrosine in the intracellular domain when a ligand is bound. Examples of the growth factor receptor family include insulin receptor, M-CSF (macrophage colony stimulating factor) receptor, EGF (epidermal growth factor) receptor, PDGF (platelet derived growth factor) receptor, FGF (fibroblast growth). factor) receptor, IGF (insulin-like growth factor) receptor, or HGF (hepatocyte growth factor) receptor.

  The TGF-β receptor family has a serine threonine kinase inside the cell membrane, and is a receptor activated by phosphorylation of serine or threonine. Examples of the TGF-β receptor family include a TGF receptor, an activin receptor, a BMP (bone morphogenic protein) receptor, and the like.

  Examples of non-kinase type receptors include cytokine receptor (type I cytokine receptor) superfamily, IFN (interferon) receptor (type II cytokine receptor) family, TNF (tumor necrosis factor) receptor (type III) A cytokine receptor) family, an immunoglobulin superfamily, or a G protein coupled receptor family.

  The cytokine receptor superfamily is a transmembrane receptor and has a characteristic amino acid sequence called a repeat of four cysteine residues or a WS box on the N-terminal side, and extracellularly fibronectin ( FN) is a receptor having a repeating sequence of a structure. Examples of the cytokine receptor superfamily include IL (interleukin) -2 receptor, IL-3 receptor, IL-4 receptor, IL-5 receptor, IL-6 receptor, IL-7 receptor, IL -9 receptor, IL-11 receptor, IL-12 receptor, IL-13 receptor, IL-15 receptor, GM-CSF (granulocyte-macrophage colony stimulating factor) receptor, CNTF (ciliary neurotrophic factor) receptor Body, EPO (erythropoietin) receptor, LIF (Leukemia inhibitory factor) receptor, OSM (oncostatin M) receptor, or G-CSF (granulocyte-colony stimulating factor) receptor. In many cases, it is composed of 2 to 3 subunits, but subunits involved in signal transduction are shared among a plurality of receptors.

  The IFN receptor family does not have a WS box, but is a receptor that is structurally similar to the cytokine receptor superfamily, such as a single-transmembrane structure and repeated cysteine residues. Examples of the IFN receptor family include IFNα receptor, IFNβ receptor, IFNγ receptor, and IL-10 receptor.

  The TNF receptor family is a receptor whose extracellular domain is composed of units rich in cysteine residues. This unit has homology among receptors of each TNF receptor family. In the intracellular domains of TNF receptor I and Fas, there is a sequence called a death domain. Examples of the TNF receptor family include TNF receptor, NGF (nerve growth factor) receptor, Fas, CD27, CD30, CD40, HVEM (Herpes virus entry mediator), LTB (Lymphotoxin β) receptor, OX40, RANK (Receptor) activator of NF-κB) or CD137, many of which are associated with cell activation and cell death.

  The immunoglobulin superfamily is a receptor having a structure having three immunoglobulin-like domains outside the cell. Examples of the immunoglobulin superfamily include IL-1 receptor or IL-18 receptor. Moreover, structurally similar Toll or Toll-like receptors can be mentioned. These include TLR1 to TLR12, and TLR4 that transmits information when endotoxin acts on cells is also a receptor of this family.

The G protein-coupled receptor (GPCR) family is a receptor that works in conjunction with G protein and has a structure that penetrates the cell membrane seven times. Examples of the GPCR family include rhodopsin receptor (Gt), catecholamine receptor [β ₁ , β ₂ , β ₃ (above, Gs), α ₁ (Gq), α ₂ (Gs, Gi, Gz, Gq)]. , Dopamine receptors [D1, D5 (above, Gs), D2 (Gi)], muscarinic receptors [m1, m3 (above, Gq), m2, m4, m5 (above, Gi, Go], adenosine receptors [ A1, A3 (or more, Gi), A2 (Gs)], PGE receptor [EP2, EP4 (or more, Gs), EP3 (Gi), EP1], Leukotriene receptor [BLT, Cys-LT1 (or more, Gi, Gq)], sphingosine 1-phosphate receptor [S1P ₁ (Gi), S1P ₂ , S1P ₃ (Gi, Gq, G ₁₃ ], lysophosphatidic acid receptor [LPA ₁ (Gi, G _{12 / 13} ), LPA ₂ , LPA ₃ (Gs, Gq)], nociceptin receptor, hGPCR44 (Gi), hGPCR4, hGPCR10, hGPCR17, hGPCR19, hGPCR39, or hGPCR48. The description in parentheses following the indicates the receptor subtype and the type of G protein α subunit that works in conjunction.

  More than 1000 sequences have already been determined as amino acid sequences (including both full-length and partial sequences) of GPCRs, and known databases such as GPCRDB [Centre for Molecular and Biomolecular Informatics (CMBI), University of Nijmegen, The Netherlands; Horn, F. et al., “GPCRDB: an information system for G protein-coupled receptors”, Nucleic Acids Res., 26 (1), 277-281, 1998]. The amino acid sequence of the human-derived GPCR shown in FIG. In the list shown in Table 1, “serial number”, “identification name in GPCRDB”, “acceptance number in GPCRDB”, “gene name”, and “description of family name” in order from the left. For example, in the GPCR with serial number 1, “5H1A_HUMAN” is the GPCRDB identifier, “P08908” in parentheses is the GPCRDB accession number, “HTR1A” is the gene name, and “Serotonin vertebrate type 1 (Serotonin "Vertebrate type 1)" is an explanation of family names. Note that “none” shown in the gene name column means that there is no gene name, and those having a plurality of gene names are also indicated by “OR”.

<Table 1>
1. 5H1A_HUMAN (P08908) HTR1A Serotonin Vertebrate type 1
2. 5H1B_HUMAN (P28222) HTR1B OR HTR1DB Serotonin Vertebrate Type 1
3. 5H1D_HUMAN (P28221) HTR1D OR HTR1DA Serotonin Vertebrate Type 1
4. 5H1E_HUMAN (P28566) HTR1E Serotonin Vertebrate Type 1
5. 5H1F_HUMAN (P30939) HTR1F OR HTR1EL Serotonin Vertebrate Type 1
6. 5H2A_HUMAN (P28223) HTR2A OR HTR2 Serotonin Vertebrate Type 2
7. 5H2B_HUMAN (P41595) HTR2B Serotonin Vertebrate Type 2
8. 5H2C_HUMAN (P28335) HTR2C OR HTR1C Serotonin Vertebrate Type 2
9. 5H4_HUMAN (Q13639) HTR4 Serotonin Vertebrate Type 4
10. 5H5A_HUMAN (P47898) HTR5A Serotonin Vertebrate Type 5
11. 5H6_HUMAN (P50406) HTR6 Serotonin Vertebrate Type 6
12. 5H7_HUMAN (P34969) HTR7 Serotonin Vertebrate Type 7
13. A1AA_HUMAN (P35348) ADRA1A OR ADRA1C Alpha Adrenoceptors type 1
14. A1AB_HUMAN (P35368) ADRA1B α-adrenergic receptor type 1
15. A1AD_HUMAN (P25100) ADRA1D OR ADRA1A α-adrenergic receptor type 1
16. A2AA_HUMAN (P08913) ADRA2A OR ADRA2R OR ADRAR α-adrenergic receptor type 2
17. A2AB_HUMAN (P18089) ADRA2B α-adrenergic receptor type 2
18. A2AC_HUMAN (P18825) ADRA2C α-adrenergic receptor type 2
19. AA1R_HUMAN (P30542) ADORA1 Adenosine type 1
20. AA2A_HUMAN (P29274) ADORA2A OR ADORA2 Adenosine Type 2
21. AA2B_HUMAN (P29275) ADORA2B Adenosine Type 2
22. AA3R_HUMAN (P33765) ADORA3 Adenosine Type 3
23. ACM1_HUMAN (P11229) CHRM1 Acetylcholine Vertebrate type 1
24. ACM2_HUMAN (P08172) CHRM2 Acetylcholine Vertebrate Type 2
25. ACM3_HUMAN (P20309) CHRM3 Acetylcholine Vertebrate Type 3
26. ACM4_HUMAN (P08173) CHRM4 Acetylcholine Vertebrate Type 4
27. ACM5_HUMAN (P08912) CHRM5 Acetylcholine Vertebrate Type 5
28. ACTR_HUMAN (Q01718) MC2R OR ACTHR Adrenocorticotropic hormone
29. ADMR_HUMAN (O15218) ADMR Adrenomedullin (G10D)
30. AG22_HUMAN (P50052) AGTR2 Angiotensin type 2
31. AG2R_HUMAN (P30556) AGTR1 OR AGTR1A OR AT2R1 Angiotensin type 1
32. AG2S_HUMAN (Q13725) AGTR1B OR AT2R1B Angiotensin type 1
33. APJ_HUMAN (P35414) AGTRL1 OR APJ APJ (APJ like)
34. B1AR_HUMAN (P08588) ADRB1 OR ADRB1R OR B1AR β-adrenergic receptor type 1
35. B2AR_HUMAN (P07550) ADRB2 OR ADRB2R OR B2AR β-adrenergic receptor type 2
36. B3AR_HUMAN (P13945) ADRB3 OR ADRB3R OR B3AR β-adrenergic receptor type 3
37. BAI1_HUMAN (O14514) BAI1 Brain-specific angiogenesis inhibitor (BAI)
38. BAI2_HUMAN (O60241) BAI2 Brain-specific angiogenesis inhibitor (BAI)
39. BAI3_HUMAN (O60242) BAI3 OR KIAA0550 Brain-specific angiogenesis inhibitor (BAI)
40. BRB1_HUMAN (P46663) BDKRB1 OR BRADYB1 Bradykinin
41. BRB2_HUMAN (P30411) BDKRB2 OR BKR2 Bradykinin
42. BRS3_HUMAN (P32247) BRS3 Bombesin
43. C3AR_HUMAN (Q16581) C3AR1 OR C3R1 OR AZ3B OR HNFAG09 C5a Anaphylatoxin
44. C3X1_HUMAN (P49238) CX3CR1 OR GPR13 C-X3-C Chemokine
45. C5AR_HUMAN (P21730) C5R1 OR C5AR C5a Anaphylatoxy
46. C5L2_HUMAN (Q9P296) GPR77 OR C5L2 C5a Anaphylatoxy
47. CALR_HUMAN (P30988) CALCR Calcitonin
48. CASR_HUMAN (P41180) CASR OR GPRC2A OR PCAR1 Extracellular calcium-sensing
49. CB1R_HUMAN (P21554) CNR1 OR CNR Cannabinoid
50. CB2R_HUMAN (P34972) CNR2 Cannabinoid
51. CCKR_HUMAN (P32238) CCKAR OR CCKRA CCK Type A
52. CCR3_HUMAN (P49682) CXCR3 OR GPR9 CXC Chemokine Type 3
53. CCR4_HUMAN (P30991) CXCR4 CXC Chemokine type 4, and chimpanzee [Pan troglodytes (Chimpanzee)]
54. CCR5_HUMAN (P32302) BLR1 OR CXCR5 CXC Chemokine Type 5
55. CCR6_HUMAN (O00574) CXCR6 OR BONZO OR STRL33 OR TYMSTR BONZO receptors (CXC6R)
56.CD97_HUMAN (P48960) CD97 EMR1
57.CGRR_HUMAN (Q16602) CALCRL OR CGRPR Calcitonin
58. CKD6_HUMAN (O00590) CCBP2 OR CMBRK9 OR CCR10 CC Chemokine Type X
59. CKR1_HUMAN (P32246) CCR1 OR CMKBR1 OR CMKR1 CC Chemokine Type 1
60. CKR2_HUMAN (P41597) CCR2 OR CMKBR2 CC Chemokine type 2
61. CKR3_HUMAN (P51677) CCR3 OR CMKBR3 CC Chemokine Type 3
62. CKR4_HUMAN (P51679) CCR4 OR CMKBR4 CC Chemokine Type 4
63. CKR5_HUMAN (P51681) CCR5 OR CMKBR5 CC Chemokine type 5
64. CKR6_HUMAN (P51684) CCR6 OR CMKBR6 OR STRL22 OR GPR29 OR CKRL3 CC Chemokine type 6
65. CKR7_HUMAN (P32248) CCR7 OR CMKBR7 OR EBI1 OR EVI1 CC Chemokine Type 7
66. CKR8_HUMAN (P51685) CCR8 OR CMKBR8 OR CKRL1 CC Chemokine type 8
67. CKR9_HUMAN (P51686) CCR9 OR CMKBR9 CC Chemokine Type 9
68. CKRA_HUMAN (P46092) GPR2 OR CCR10 CC Chemokine type 10
69. CKRB_HUMAN (Q9NPB9) CCRL1 OR CCR11 OR CCBP2 OR VSHK1 CC Chemokine type 11
70. CLR1_HUMAN (Q9NYQ6) CELSR1 OR CDHF9 OR FMI2 Cadherin EGF LAG (CELSR)
71. CLR2_HUMAN (Q9HCU4) CELSR2 OR CDHF10 OR EGFL2 OR MEGF3 OR KIAA0279 Cadherin EGF LAG (CELSR)
72. CLR3_HUMAN (Q9NYQ7) CELSR3 OR CDHF11 OR FMI1 OR EGFL1 OR MEGF2 Cadherin EGF LAG (CELSR)
73. CLT1_HUMAN (Q9Y271) CYSLTR1 OR CYSLT1 Cysteinyl leukotriene
74. CLT2_HUMAN (Q9NS75) CYSLTR2 OR CYSLT2 OR CYSLT2R Cysteinyl leukotriene
75. CML1_HUMAN (Q99788) CMKLR1 OR DEZ OR CHEMR23 Chemokine receptor-like 1
76. CML2_HUMAN (Q99527) GPR30 OR CMKRL2 OR DRY12 OR CEPR Chemokine receptor-like 2
77. CRF1_HUMAN (P34998) CRHR1 OR CRHR OR CRFR Corticotropin releasing factor
78. CRF2_HUMAN (Q13324) CRHR2 OR CRF2R OR CRH2R Corticotropin releasing factor
79. CRF_HUMAN (P06850) CRH fragment
80. CXC1_HUMAN (P46094) XCR1 OR CCXCR1 OR GPR5 XC Chemokine
81. D2DR_HUMAN (P14416) DRD2 Dopamine Vertebrate type 2
82. D3DR_HUMAN (P35462) DRD3 Dopamine Vertebrate Type 3
83. D4DR_HUMAN (P21917) DRD4 Dopamine Vertebrate Type 4
84. DADR_HUMAN (P21728) DRD1 Dopamine Vertebrate Type 1
85. DBDR_HUMAN (P21918) DRD5 OR DRD1B Dopamine Vertebrate Type 1
86. DUFF_HUMAN (Q16570) FY OR DARC OR GPD Estimated / Unclassified Class A GPCRs (Putative / unclassified Class A GPCRs)
87. EBI2_HUMAN (P32249) EBI2 EBV-induced
88. EDG1_HUMAN (P21453) EDG1 Lysosphingolipid & LPA (EDG)
89. EDG2_HUMAN (Q92633) EDG2 Lysosphingolipid & LPA (EDG)
90. EDG3_HUMAN (Q99500) EDG3 Lysosphingolipid & LPA (EDG)
91. EDG4_HUMAN (Q9HBW0) EDG4 Lysosphingolipid & LPA (EDG)
92.EDG5_HUMAN (O95136) EDG5 Lysosphingolipid & LPA (EDG)
93. EDG7_HUMAN (Q9UBY5) EDG7 Rhizosphingolipid & LPA (EDG)
94. EMR1_HUMAN (Q14246) EMR1 EMR1
95. ET1R_HUMAN (P25101) EDNRA OR ETRA OR ETA Endothelin
96. ETB2_HUMAN (O60883) ETBRLP2 GPR37 / endothelin B-like
97. ETBR_HUMAN (P24530) EDNRB OR ETRB Endothelin
98. FK79_HUMAN (Q9BXC1) FKSG79 Purinoceptor Other
99.FML2_HUMAN (P25089) FPRL2 OR FPRH1 Fmet-leu-phe
100. FMLR_HUMAN (P21462) FPR1 Fmet-leu-phe
101. FSHR_HUMAN (P23945) FSHR Follicle stimulating hormone
102. FZ10_HUMAN (Q9ULW2) FZD10 Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
103. FZD1_HUMAN (Q9UP38) FZD1 Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
104. FZD2_HUMAN (Q14332) FZD2 Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
105. FZD3_HUMAN (Q9NPG1) FZD3 Frizzled Group B (Fz 3 & 6)
106. FZD4_HUMAN (Q9ULV1) FZD4 Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
107. FZD5_HUMAN (Q13467) FZD5 OR HFZ5 Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
108. FZD6_HUMAN (O60353) FZD6 Frizzled Group B (Fz 3 & 6)
109. FZD7_HUMAN (O75084) FZD7 Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
110. FZD8_HUMAN (Q9H461) FZD8 Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
111. FZD9_HUMAN (O00144) FZD9 OR FZD3 Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
112. G101_HUMAN (Q96P66) GPR101 Estimated / Unclassified Class A GPCRs
113. G103_HUMAN (Q96P65) GPR103 Estimated / Unclassified Class A GPCRs
114. G2A_HUMAN (Q9UNW8) G2A Estimated / Unclassified Class A GPCRs
115. GALR_HUMAN (P47211) GALR1 OR GALNR1 OR GALNR Galanin
116. GALS_HUMAN (O43603) GALR2 OR GALNR2 Galanin
117. GALT_HUMAN (O60755) GALR3 OR GALNR3 Galanin
118. GASR_HUMAN (P32239) CCKBR OR CCKRB CCK Type B
119. GBR1_HUMAN (Q9UBS5) GABBR1 GABA-B Subtype 1
120. GBR2_HUMAN (O75899) GPR51 OR GABBR2 GABA-B Subtype 2
121.GHSR_HUMAN (Q92847) GHSR Growth hormone secretagogue
122.GIPR_HUMAN (P48546) GIPR Gastric inhibitory peptide
123. GLP1_HUMAN (P43220) GLP1R Glucagon
124. GLP2_HUMAN (O95838) GLP2R Glucagon
125. GLR_HUMAN (P47871) GCGR Glucagon
126.GP10_HUMAN (P49683) GPR10 OR GR3 GPR
127.GP12_HUMAN (P47775) GPR12 GPR
128. GP15_HUMAN (P49685) GPR15 GPR
129. GP17_HUMAN (Q13304) GPR17 GPR
130. GP18_HUMAN (Q14330) GPR18 OR GPCRW Estimated / Unclassified Class A GPCRs
131.GP19_HUMAN (Q15760) GPR19 GPR
132.GP20_HUMAN (Q99678) GPR20 GPR
133. GP21_HUMAN (Q99679) GPR21 GPR
134.GP22_HUMAN (Q99680) GPR22 GPR
135. GP25_HUMAN (O00155) GPR25 GPR
136. GP26_HUMAN (Q8NDV2) GPR26 Estimated / Unclassified Class A GPCRs
137. GP27_HUMAN (Q9NS67) GPR27 OR SREB1 SREB
138. GP31_HUMAN (O00270) GPR31 GPR
139. GP32_HUMAN (O75388) GPR32 Chemokine Receptor-like 1
140. GP34_HUMAN (Q9UPC5) GPR34 Estimated / Unclassified A GPCRs
141. GP35_HUMAN (Q9HC97) GPR35 Purine Receptor etc.
142. GP37_HUMAN (O15354) GPR37 GPR37 / Endothelin B-like
143. GP39_HUMAN (O43194) GPR39 Estimated / Unclassified Class A GPCRs
144. GP40_HUMAN (O14842) GPR40 GP40 (GP40 like)
145.GP41_HUMAN (O14843) GPR41 GP40
146. GP42_HUMAN (O15529) GPR42 GP40
147. GP43_HUMAN (O15552) GPR43 GP40
148. GP44_HUMAN (Q9Y5Y4) GPR44 OR CRTH2 OR DL1R Chemokine receptor-like 1
149. GP45_HUMAN (Q9Y5Y3) GPR45 GPR45
150. GP52_HUMAN (Q9Y2T5) GPR52 GPR
151. GP57_HUMAN (Q9P1P4) GPR57 Trace amine
152. GP58_HUMAN (Q9P1P5) GPR58 Trace amine
153. GP61_HUMAN (Q9BZJ8) GPR61 OR BALGR Estimated / Unclassified Class A GPCRs
154. GP62_HUMAN (Q9BZJ7) GPR62 Estimated / Unclassified Class A GPCRs
155. GP63_HUMAN (Q9BZJ6) GPR63 OR PSP24B GPR45
156. GP78_HUMAN (Q96P69) GPR78 Estimated / Unclassified Class A GPCRs
157. GP80_HUMAN (Q96P68) GPR80 OR GPR99 Estimated / Unclassified Class A GPCRs
158. GP81_HUMAN (Q9BXC0) GPR81 OR FKSG80 Estimated / Unclassified Class A GPCRs
159. GP82_HUMAN (Q96P67) GPR82 Estimated / Unclassified Class A GPCRs
160. GP83_HUMAN (Q9NYM4) GPR83 OR GPR72 OR KIAA1540 Neuropeptide Y other
161. GP86_HUMAN (Q9BPV8) GPR86 OR GPR94 OR FKSG77 Platelet ADP & KI01 receptors
162. GP87_HUMAN (Q9BY21) GPR87 OR GPR95 OR FKSG78 Platelet ADP & KI01 Receptors
163. GP92_HUMAN (Q9H1C0) GPR92 OR GPR93 Purine receptor etc.
164. GPR1_HUMAN (P46091) GPR1 GPR
165. GPR3_HUMAN (P46089) GPR3 OR ACCA GPR
166. GPR4_HUMAN (P46093) GPR4 OR GPR19 GPR
167. GPR6_HUMAN (P46095) GPR6 GPR
168. GPR7_HUMAN (P48145) GPR7 GPR
169. GPR8_HUMAN (P48146) GPR8 GPR
170. GRFR_HUMAN (Q02643) GHRHR Growth hormone-releasing hormone
171. GRHR_HUMAN (P30968) RHR Gonadotropin-releasing hormone type I
172.GRPR_HUMAN (P30550) GRPR Bombesin
173. GRR2_HUMAN (Q96P88) RHR2 Gonadotropin releasing hormone type II
174. H963_HUMAN (O14626) H963 Estimated / Unclassified Class A GPCRs
175. HH1R_HUMAN (P35367) HRH1 Histamine Type 1
176. HH2R_HUMAN (P25021) HRH2 Histamine Type 2
177. HH3R_HUMAN (Q9Y5N1) HRH3 OR GPCR97 Histamine Type 3
178. HH4R_HUMAN (Q9H3N8) HRH4 Histamine Type 4
179. HM74_HUMAN (P49019) HM74 Estimated / Unclassified Class A GPCRs
180. IL8A_HUMAN (P25024) IL8RA OR CXCR1 Interleukin-8 Type A
181. IL8B_HUMAN (P25025) IL8RB OR CXCR2 Interleukin-8 Type B
182. L4R1_HUMAN (Q15722) LTB4R OR P2RY7 OR GPR16 OR CMKRL1 OR BLT OR BLT1 Leukotriene B4 receptor BLT1
183. L4R2_HUMAN (Q9NPC1) LTB4R2 OR BLT2R OR BLTR2 Leukotriene B4 receptor BLT2
184. LGR4_HUMAN (Q9BXB1) GPR48 OR LGR4 LGR-like [hormone receptors]
185. LGR5_HUMAN (O75473) GPR49 OR LGR5 OR GPR67 LGR-like (hormone receptors)
186. LGR6_HUMAN (Q9HBX8) LGR6 LGR-like (hormone receptors)
187. LGR7_HUMAN (Q9HBX9) LGR7 LGR-like (hormone receptors)
188. LGR8_HUMAN (Q8WXD0) LGR8 OR GREAT LGR-like (hormone receptors)
189. LSHR_HUMAN (P22888) LHCGR OR LHRHR OR LCGR Lutropin-choriogonadotropic hormone
190. MAS_HUMAN (P04201) MAS1 OR MAS Mas Proto-oncogene
191. MC3R_HUMAN (P41968) MC3R Melanocortin hormone
192. MC4R_HUMAN (P32245) MC4R Melanocortin hormone
193. MC5R_HUMAN (P33032) MC5R Melanocortin hormone
194. MCR1_HUMAN (Q99705) GPR24 OR MCHR1 OR SLC1 Melanin-concentrating hormone receptors
195. MCR2_HUMAN (Q969V1) GPR145 OR MCHR2 OR SLT Melanin-concentrating hormone receptors
196. MGR1_HUMAN (Q13255) GRM1 OR GPRC1A OR MGLUR1 Metabotropic glutamate group I
197. MGR2_HUMAN (Q14416) GRM2 OR GPRC1B OR MGLUR2 Metabolic Glutamate Group II
198. MGR3_HUMAN (Q14832) GRM3 OR GPRC1C OR MGLUR3 Metabolic Glutamate Group II
199. MGR4_HUMAN (Q14833) GRM4 OR GPRC1D OR MGLUR4 Metabotropic Glutamate Group III
200. MGR5_HUMAN (P41594) GRM5 OR GPRC1E OR MGLUR5 Metabotropic Glutamate Group I
201. MGR6_HUMAN (O15303) GRM6 OR GPRC1F OR MGLUR6 Metabotropic Glutamate Group III
202. MGR7_HUMAN (Q14831) GRM7 OR GPRC1G OR MGLUR7 Metabotropic Glutamate Group III
203. MGR8_HUMAN (O00222) GRM8 OR GPRC1H OR MGLUR8 Metabotropic Glutamate Group III
204. ML1A_HUMAN (P48039) MTNR1A Melatonin
205. ML1B_HUMAN (P49286) MTNR1B Melatonin
206. ML1X_HUMAN (Q13585) GPR50 Melatonin
207. MRGF_HUMAN (Q96AM1) MRGF Estimated / Unclassified Class A GPCRs
208. MRG_HUMAN (P35410) MAS1L OR MRG Mas-related receptors (MRGs)
209. MSHR_HUMAN (Q01726) MC1R OR MSHR Melanocyte stimulating hormone
210. MTLR_HUMAN (O43193) GPR38 OR MTLR1 OR MTLR Growth hormone secretagogue like
211. NFF1_HUMAN (Q9GZQ6) NPFF1 Neuropeptide FF
212. NFF2_HUMAN (Q9Y5X5) GPR74 OR NPGPR OR NPFF2 Neuropeptide FF
213. NK1R_HUMAN (P25103) TACR1 OR TAC1R OR NK1R Substance P (NK1)
214. NK2R_HUMAN (P21452) TACR2 OR TAC2R OR NK2R Substance K (NK2)
215. NK3R_HUMAN (P29371) TACR3 OR TAC3R OR NK3R Neuromedin K (Neuromedin K) (NK3)
216. NMBR_HUMAN (P28336) NMBR Bombesin
217. NTR1_HUMAN (P30989) NTSR1 OR NTRR Neurotensin
218. NTR2_HUMAN (O95665) NTSR2 Neurotensin
219. NY1R_HUMAN (P25929) NPY1R OR NPYR OR NPYY1 Neuropeptide Y type 1
220. NY2R_HUMAN (P49146) NPY2R Neuropeptide Y type 2
221. NY4R_HUMAN (P50391) PPYR1 OR NPY4R Neuropeptide Y type 4
222. NY5R_HUMAN (Q15761) NPY5R OR NPYR5 Neuropeptide Y type 5
223. O00325 (O00325) none Prostaglandin E2 Subtype EP3
224. O00326 (O00326) none Prostaglandin E2 Subtype EP3
225. O00406 (O00406) HE6 Estimated / Unclassified Class B GPCRs
226. O00421 (O00421) CCR6 OR CKRX CC Chemokine other
227. O00718 (O00718) none EMR1
228. O14694 (O14694) CCR5 fragment
229. O14804 (O14804) none Trace amine
230. O15538 (P51681) CCR5 fragment
231.O1A1_HUMAN (Q9P1Q5) OR1A1 Olfactory II fam 1 / MOR125-138,156
232.O1A2_HUMAN (Q9Y585) OR1A2 Olfactory II fam 1 / MOR125-138,156
233. O1B1_HUMAN (Q8NGR6) OR1B1 Olfactory II fam 1 / MOR125-138,156
234. O1C1_HUMAN (Q15619) OR1C1 Olfactory II fam 1 / MOR125-138,156
235. O1D2_HUMAN (P34982) OR1D2 OR OLFR1 Olfactory II fam 1 / MOR125-138,156
236. O1D4_HUMAN (P47884) OR1D4 Olfactory II fam 1 / MOR125-138,156
237. O1D5_HUMAN (P58170) OR1D5 Olfactory II fam 1 / MOR125-138,156
238. O1E1_HUMAN (P30953) OR1E1 Olfactory II fam 1 / MOR125-138,156
239. O1E2_HUMAN (P47887) OR1E2 Olfactory II fam 1 / MOR125-138,156
240. O1E5_HUMAN (Q9UM60) OR1E5 fragment
241.O1E6_HUMAN (O43884) OR1E6 fragment
242.O1F1_HUMAN (O43749) OR1F1 OR OLFMF Olfactory II fam 1 / MOR125-138,156
243. O1F2_HUMAN (Q96R84) OR1F2 Olfactory II fam 1 / MOR125-138,156
244.O1FA_HUMAN (O43882) OR1F10 fragment
245. O1FC_HUMAN (Q8NHA8) OR1F12 Olfactory II fam 1 / MOR125-138,156
246. O1G1_HUMAN (P47890) OR1G1 Olfactory II fam 1 / MOR125-138,156
247. O1I1_HUMAN (O60431) OR1I1 Olfactory II fam 1 / MOR125-138,156
248. O1J1_HUMAN (Q8NGS3) OR1J1 Olfactory II fam 1 / MOR125-138,156
249. O1J2_HUMAN (Q8NGS2) OR1J2 Olfactory II fam 1 / MOR125-138,156
250. O1J4_HUMAN (Q8NGS1) OR1J4 Olfactory II fam 1 / MOR125-138,156
251. O1J5_HUMAN (Q9NZP1) OR1J5 fragment
252. O1K1_HUMAN (Q8NGR3) OR1K1 Olfactory II fam 1 / MOR125-138,156
253. O1L1_HUMAN (Q8NH94) OR1L1 OR OR1L2 Olfactory II fam 1 / MOR125-138,156
254. O1L3_HUMAN (Q8NH93) OR1L3 Olfactory II fam 1 / MOR125-138,156
255. O1L4_HUMAN (Q8NGR5) OR1L4 OR OR1L5 Olfactory II fam 1 / MOR125-138,156
256. O1L6_HUMAN (Q8NGR2) OR1L6 OR OR1L7 Olfactory II fam 1 / MOR125-138,156
257. O1L8_HUMAN (Q8NGR8) OR1L8 Olfactory II fam 1 / MOR125-138,156
258. O1M1_HUMAN (Q8NGA1) OR1M1 Olfactory II fam 1 / MOR125-138,156
259. O1N1_HUMAN (O43870) OR1N1 fragment
260. O1N2_HUMAN (Q8NGR9) OR1N2 Olfactory II fam 1 / MOR125-138,156
261.O1N3_HUMAN (Q8NGS0) OR1N3 Olfactory II fam 1 / MOR125-138,156
262. O1Q1_HUMAN (Q15612) OR1Q1 OR OR1Q2 OR OR1Q3 Olfactory II fam 1 / MOR125-138,156
263.O1S1_HUMAN (Q8NH92) OR1S1 Olfactory II fam 1 / MOR125-138,156
264. O1S2_HUMAN (Q8NGQ3) OR1S2 Olfactory II fam 1 / MOR125-138,156
265. O2A1_HUMAN (Q8NGT9) OR2A1 Olfactory II fam 2 / MOR256-262,270-285
266. O2A4_HUMAN (O95047) OR2A4 OR OR2A10 Olfactory II fam 2 / MOR256-262,270-285
267. O2A5_HUMAN (Q96R48) OR2A5 OR OR2A8 fragment
268. O2A6_HUMAN (Q96R47) OR2A6 fragment
269. O2A7_HUMAN (Q96R45) OR2A7 Olfactory II fam 2 / MOR256-262,270-285
270. O2AC_HUMAN (Q8NGT7) OR2A12 Olfactory II fam 2 / MOR256-262,270-285
271. O2AE_HUMAN (Q8NHA4) OR2AE1 OR OR2AE2 Olfactory II fam 2 / MOR256-262,270-285
272. O2AG_HUMAN (Q9H205) OR2AG1 Olfactory II fam 2 / MOR256-262,270-285
273.O2AK_HUMAN (Q8NG84) OR2AK2 Olfactory II fam 2 / MOR256-262,270-285
274. O2B1_HUMAN (O43883) OR2B1 OR OR2B5 fragment
275. O2B2_HUMAN (Q9GZK3) OR2B2 OR OR2B9 Olfactory II fam 2 / MOR256-262,270-285
276. O2B3_HUMAN (O76000) OR2B3 Olfactory II fam 2 / MOR256-262,270-285
277. O2B6_HUMAN (P58173) OR2B6 Olfactory II fam 2 / MOR256-262,270-285
278. O2C1_HUMAN (O95371) OR2C1 Olfactory II fam 2 / MOR256-262,270-285
279. O2C3_HUMAN (Q8N628) OR2C3 OR OR2C4 Olfactory II fam 2 / MOR256-262,270-285
280. O2D2_HUMAN (Q9H210) OR2D2 Olfactory II fam 2 / MOR256-262,270-285
281. O2D3_HUMAN (Q8NGH3) OR2D3 Olfactory II fam 2 / MOR256-262,270-285
282. O2F1_HUMAN (Q13607) OR2F1 OR OLF3 Olfactory II fam 2 / MOR256-262,270-285
283. O2F2_HUMAN (O95006) OR2F2 Olfactory II fam 2 / MOR256-262,270-285
284. O2G2_HUMAN (Q8NGZ5) OR2G2 Olfactory II fam 2 / MOR256-262,270-285
285. O2G3_HUMAN (Q8NGZ4) OR2G3 Olfactory II fam 2 / MOR256-262,270-285
286. O2H1_HUMAN (Q9GZK4) OR2H1 OR OR2H8 Olfactory II fam 2 / MOR256-262,270-285
287. O2H2_HUMAN (O95918) OR2H2 Olfactory II fam 2 / MOR256-262,270-285
288. O2H3_HUMAN (Q15062) OR2H3 OR OLFR2 OR FAT11 Olfactory II fam 2 / MOR256-262,270-285
289. O2H7_HUMAN (O43628) OR2H7 fragment
290. O2I2_HUMAN (Q8NGU4) OR2I2 Olfactory II fam 2 / MOR256-262,270-285
291. O2J2_HUMAN (O76002) OR2J2 Olfactory II fam 2 / MOR256-262,270-285
292. O2J3_HUMAN (O76001) OR2J3 Olfactory II fam 2 / MOR256-262,270-285
293. O2K2_HUMAN (Q8NGT1) OR2K2 Olfactory II fam 2 / MOR256-262,270-285
294. O2L2_HUMAN (Q8NH16) OR2L2 Olfactory II fam 2 / MOR256-262,270-285
295. O2L8_HUMAN (Q8NGY9) OR2L8 Olfactory II fam 2 / MOR256-262,270-285
296. O2LB_HUMAN (Q8NG80) OR2L11 Olfactory II fam 2 / MOR256-262,270-285
297. O2LD_HUMAN (Q8N349) OR2L14 OR OR2L13 Olfactory II fam 2 / MOR256-262,270-285
298. O2M1_HUMAN (Q96R29) OR2M1 fragment
299. O2M2_HUMAN (Q96R28) OR2M2 fragment
300. O2M4_HUMAN (Q96R27) OR2M4 fragment
301. O2M6_HUMAN (Q8NG83) OR2M6 Olfactory II fam 2 / MOR256-262,270-285
302. O2M7_HUMAN (Q8NG81) OR2M7 Olfactory II fam 2 / MOR256-262,270-285
303. O2S2_HUMAN (Q9NQN1) OR2S2 Olfactory II fam 2 / MOR256-262,270-285
304. O2T1_HUMAN (O43869) OR2T1 Olfactory II fam 2 / MOR256-262,270-285
305. O2T2_HUMAN (Q8NGX2) OR2T2 Olfactory II fam 2 / MOR256-262,270-285
306. O2T3_HUMAN (Q8NH03) OR2T3 fragment
307. O2T4_HUMAN (Q8NH00) OR2T4 Olfactory II fam 2 / MOR256-262,270-285
308. O2T5_HUMAN (Q8NH02) OR2T5 Olfactory II fam 2 / MOR256-262,270-285
309. O2TB_HUMAN (Q8NH01) OR2T11 Olfactory II fam 2 / MOR256-262,270-285
310. O2V2_HUMAN (Q96R30) OR2V2 Olfactory II fam 2 / MOR256-262,270-285
311. O2V3_HUMAN (Q8NGV1) OR2V3 Olfactory II fam 2 / MOR256-262,270-285
312. O2W1_HUMAN (Q9Y3N9) OR2W1 Olfactory II fam 2 / MOR256-262,270-285
313. O2Y1_HUMAN (Q8NGV0) OR2Y1 Olfactory II fam 2 / MOR256-262,270-285
314. O2Z1_HUMAN (Q8NG97) OR2Z1 OR OR2Z2 Olfactory II fam 2 / MOR256-262,270-285
315. O3A1_HUMAN (P47881) OR3A1 OR OLFRA03 Olfactory II fam 3 / MOR255
316. O3A2_HUMAN (P47893) OR3A2 OR OLFRA04 Olfactory II fam 3 / MOR255
317. O3A3_HUMAN (P47888) OR3A3 Olfactory II fam 3 / MOR255
318. O3A4_HUMAN (P47883) OR3A4 Olfactory II fam 3 / MOR255
319. O43192 (O43192) none Vasopressin
320. O43200 (O43200) TSHR fragment
321.O43441 (O43441) ETA fragment
322.O43624 (O43624) OLFR 42A fragment
323.O43625 (O43625) OLFR 42A fragment
324.O43626 (O43626) OLFR 42B fragment
325.O43627 (O43627) OLFR 42B fragment
326.O43662 (O43662) OLFR42A Fragment
327. O43664 (Q9HB89) none Neuromedin U
328. O43789 (O43789) none fragment
329. O43871 (O43871) OR16-36 fragment
330.O43872 (O43872) OR16-37 fragment
331. O43873 (O43873) OR16-88 fragment
332.O43874 (O43874) OR16-89 fragment
333. O43875 (O43875) OR16-90 fragment
334. O43876 (O43876) OR17-130 fragment
335. O43878 (O43878) OR17-137 fragment
336. O43879 (O43879) OR17-15 fragment
337. O43880 (O43880) OR17-16 fragment
338. O43886 (O43886) OR7-139 fragment
339.O43887 (O43887) OR7-140 Fragment
340. O43888 (O43888) OR7-141 Fragment
341.O43898 (O43898) none GPR45
342. O4A4_HUMAN (Q8NGN8) OR4A4 Olfactory II fam 4 / MOR225-248
343. O4A5_HUMAN (Q8NH83) OR4A5 Olfactory II fam 4 / MOR225-248
344. O4AF_HUMAN (Q8NGL6) OR4A15 Olfactory II fam 4 / MOR225-248
345.O4AG_HUMAN (Q8NH70) OR4A16 Olfactory II fam 4 / MOR225-248
346. O4B1_HUMAN (Q8NGF8) OR4B1 Olfactory II fam 4 / MOR225-248
347. O4C3_HUMAN (Q8NH37) OR4C3 Olfactory II fam 4 / MOR225-248
348. O4C6_HUMAN (Q8NH72) OR4C6 Olfactory II fam 4 / MOR225-248
349. O4CC_HUMAN (Q96R67) OR4C12 Olfactory II fam 4 / MOR225-248
350. O4CD_HUMAN (Q8NGP0) OR4C13 Olfactory II fam 4 / MOR225-248
351. O4CF_HUMAN (Q8NGM1) OR4C15 Olfactory II fam 4 / MOR225-248
352. O4CG_HUMAN (Q8NGL9) OR4C16 Olfactory II fam 4 / MOR225-248
353. O4D1_HUMAN (Q15615) OR4D1 Olfactory II fam 4 / MOR225-248
354. O4D2_HUMAN (P58180) OR4D2 Olfactory II fam 4 / MOR225-248
355. O4D5_HUMAN (Q8NGN0) OR4D5 Olfactory II fam 4 / MOR225-248
356. O4D6_HUMAN (Q8NGJ1) OR4D6 Olfactory II fam 4 / MOR225-248
357. O4D9_HUMAN (Q8NGE8) OR4D9 Olfactory II fam 4 / MOR225-248
358. O4E2_HUMAN (Q8NGC2) OR4E2 Olfactory II fam 4 / MOR225-248
359. O4F3_HUMAN (O95013) OR4F3 Olfactory II fam 4 / MOR225-248
360. O4F4_HUMAN (Q96R69) OR4F4 Olfactory II fam 4 / MOR225-248
361.O4F5_HUMAN (Q8NH21) OR4F5 Olfactory II fam 4 / MOR225-248
362.O4F6_HUMAN (Q8NGB9) OR4F6 OR OR4F12 Olfactory II fam 4 / MOR225-248
363. O4FE_HUMAN (Q8NGB7) OR4F14 Olfactory II fam 4 / MOR225-248
364. O4FF_HUMAN (Q8NGB8) OR4F15 Olfactory II fam 4 / MOR225-248
365. O4FH_HUMAN (Q8NGA8) OR4F17 OR OR4F19 Olfactory II fam 4 / MOR225-248
366. O4HC_HUMAN (Q8NG74) OR4H12 fragment
367. O4K1_HUMAN (Q8NGD4) OR4K1 Olfactory II fam 4 / MOR225-248
368. O4K2_HUMAN (Q8NGD2) OR4K2 Olfactory II fam 4 / MOR225-248
369. O4K3_HUMAN (Q96R72) OR4K3 Olfactory II fam 4 / MOR225-248
370. O4K5_HUMAN (Q8NGD3) OR4K5 Olfactory II fam 4 / MOR225-248
371. O4KD_HUMAN (Q8NH42) OR4K13 Olfactory II fam 4 / MOR225-248
372. O4KF_HUMAN (Q8NH41) OR4K15 Olfactory II fam 4 / MOR225-248
373. O4KH_HUMAN (Q8NGC6) OR4K17 Olfactory II fam 4 / MOR225-248
374. O4L1_HUMAN (Q8NH43) OR4L1 Olfactory II fam 4 / MOR225-248
375. O4M1_HUMAN (Q8NGD0) OR4M1 Olfactory II fam 4 / MOR225-248
376. O4M2_HUMAN (Q8NGB6) OR4M2 Olfactory II fam 4 / MOR225-248
377. O4N2_HUMAN (Q8NGD1) OR4N2 Olfactory II fam 4 / MOR225-248
378. O4N4_HUMAN (Q8N0Y3) OR4N4 Olfactory II fam 4 / MOR225-248
379. O4N5_HUMAN (Q8IXE1) OR4N5 Olfactory II fam 4 / MOR225-248
380. O4P4_HUMAN (Q8NGL7) OR4P4 Olfactory II fam 4 / MOR225-248
381. O4Q4_HUMAN (Q8NH05) OR4Q4 Olfactory II fam 4 / MOR225-248
382. O4S1_HUMAN (Q8NGB4) OR4S1 Olfactory II fam 4 / MOR225-248
383. O4S2_HUMAN (Q8NH73) OR4S2 Olfactory II fam 4 / MOR225-248
384. O4X1_HUMAN (Q8NH49) OR4X1 Olfactory II fam 4 / MOR225-248
385. O4X2_HUMAN (Q8NGF9) OR4X2 Olfactory II fam 4 / MOR225-248
386. O5A1_HUMAN (Q8NGJ0) OR5A1 Olfactory II fam 5 / MOR172-224,249,254
387. O5A2_HUMAN (Q8NGI9) OR5A2 Olfactory II fam 5 / MOR172-224,249,254
388. O5AC_HUMAN (Q9NZP5) OR5AC2 fragment
389. O5AK_HUMAN (Q8NH90) OR5AK2 Olfactory II fam 5 / MOR172-224,249,254
390. O5AN_HUMAN (Q8NGI8) OR5AN1 Olfactory II fam 5 / MOR172-224,249,254
391. O5AP_HUMAN (Q8NGF4) OR5AP2 Olfactory II fam 5 / MOR172-224,249,254
392.O5AR_HUMAN (Q8NGP9) OR5AR1 Olfactory II fam 5 / MOR172-224,249,254
393. O5AS_HUMAN (Q8N127) OR5AS1 Olfactory II fam 5 / MOR172-224,249,254
394. O5AT_HUMAN (Q8NHC5) OR5AT1 Olfactory II fam 5 / MOR172-224,249,254
395. O5AU_HUMAN (Q8NGC0) OR5AU1 Olfactory II fam 5 / MOR172-224,249,254
396. O5AV_HUMAN (Q8NHC6) OR5AV1 Olfactory II fam 5 / MOR172-224,249,254
397. O5AY_HUMAN (Q8NGZ2) OR5AY1 Olfactory II fam 5 / MOR172-224,249,254
398. O5B2_HUMAN (Q96R09) OR5B2 Olfactory II fam 5 / MOR172-224,249,254
399. O5BD_HUMAN (Q8NH48) OR5B13 Olfactory II fam 5 / MOR172-224,249,254
400. O5BF_HUMAN (Q8NHC7) OR5BF1 Olfactory II fam 5 / MOR172-224,249,254
401.O5BG_HUMAN (Q96R08) OR5B16 Olfactory II fam 5 / MOR172-224,249,254
402. O5BH_HUMAN (Q8NGF7) OR5B17 Olfactory II fam 5 / MOR172-224,249,254
403. O5C1_HUMAN (Q8NGR4) OR5C1 Olfactory II fam 5 / MOR172-224,249,254
404. O5DD_HUMAN (Q8NGL4) OR5D13 Olfactory II fam 5 / MOR172-224,249,254
405. O5DE_HUMAN (Q8NGL3) OR5D14 Olfactory II fam 5 / MOR172-224,249,254
406. O5DG_HUMAN (Q8NGK9) OR5D16 Olfactory II fam 5 / MOR172-224,249,254
407. O5DI_HUMAN (Q8NGL1) OR5D18 Olfactory II fam 5 / MOR172-224,249,254
408. O5F1_HUMAN (O95221) OR5F1 Olfactory II fam 5 / MOR172-224,249,254
409. O5H2_HUMAN (Q8NGV7) OR5H2 Olfactory II fam 5 / MOR172-224,249,254
410. O5H6_HUMAN (Q8NGV6) OR5H6 Olfactory II fam 5 / MOR172-224,249,254
411. O5I1_HUMAN (Q13606) OR5I1 OR OLF1 Olfactory II fam 5 / MOR172-224,249,254
412. O5J2_HUMAN (Q8NH18) OR5J2 Olfactory II fam 5 / MOR172-224,249,254
413. O5K1_HUMAN (Q8NHB7) OR5K1 Olfactory II fam 5 / MOR172-224,249,254
414. O5K2_HUMAN (Q8NHB8) OR5K2 Olfactory II fam 5 / MOR172-224,249,254
415. O5K3_HUMAN (Q8NH89) OR5AK3 Olfactory II fam 5 / MOR172-224,249,254
416. O5L1_HUMAN (Q8NGL2) OR5L1 Olfactory II fam 5 / MOR172-224,249,254
417.O5L2_HUMAN (Q8NGL0) OR5L2 Olfactory II fam 5 / MOR172-224,249,254
418. O5M1_HUMAN (Q8NGP8) OR5M1 Olfactory II fam 5 / MOR172-224,249,254
419. O5M3_HUMAN (Q8NGP4) OR5M3 Olfactory II fam 5 / MOR172-224,249,254
420. O5M8_HUMAN (Q8NGP6) OR5M8 Olfactory II fam 5 / MOR172-224,249,254
421.O5M9_HUMAN (Q8NGP3) OR5M9 Olfactory II fam 5 / MOR172-224,249,254
422. O5MB_HUMAN (Q96RB7) OR5M11 Olfactory II fam 5 / MOR172-224,249,254
423.O5P2_HUMAN (Q8WZ92) OR5P2 Olfactory II fam 5 / MOR172-224,249,254
424. O5P3_HUMAN (Q8WZ94) OR5P3 Olfactory II fam 5 / MOR172-224,249,254
425. O5T2_HUMAN (Q8NGG2) OR5T2 Olfactory II fam 5 / MOR172-224,249,254
426. O5T3_HUMAN (Q8NGG3) OR5T3 Olfactory II fam 5 / MOR172-224,249,254
427. O5U1_HUMAN (Q9UGF5) OR5U1 Olfactory II fam 5 / MOR172-224,249,254
428. O5V1_HUMAN (Q9UGF6) OR5V1 Olfactory II fam 5 / MOR172-224,249,254
429.O60411 (O60411) none fragment
430. O60451 (O60451) none α-adrenergic receptor type 1
431. O6A1_HUMAN (O95222) OR6A1 Olfactory II fam 6 / MOR103-105,107-119
432.O6B1_HUMAN (O95007) OR6B1 Olfactory II fam 6 / MOR103-105,107-119
433. O6B3_HUMAN (Q8NGW1) OR6B3 Olfactory II fam 6 / MOR103-105,107-119
434. O6C1_HUMAN (Q96RD1) OR6C1 fragment
435. O6C2_HUMAN (Q9NZP2) OR6C2 fragment
436. O6C3_HUMAN (Q9NZP0) OR6C3 fragment
437. O6C4_HUMAN (Q8NGE1) OR6C4 Olfactory II fam 6 / MOR103-105,107-119
438. O6F1_HUMAN (Q8NGZ6) OR6F1 Olfactory II fam 6 / MOR103-105,107-119
439. O6J2_HUMAN (Q8NGC5) OR6J2 Olfactory II fam 6 / MOR103-105,107-119
440. O6K2_HUMAN (Q8NGY2) OR6K2 Olfactory II fam 6 / MOR103-105,107-119
441. O6K3_HUMAN (Q8NGY3) OR6K3 Olfactory II fam 6 / MOR103-105,107-119
442. O6K6_HUMAN (Q8NGW6) OR6K6 Olfactory II fam 6 / MOR103-105,107-119
443. O6M1_HUMAN (Q8NGM8) OR6M1 Olfactory II fam 6 / MOR103-105,107-119
444. O6N1_HUMAN (Q8NGY5) OR6N1 Olfactory II fam 6 / MOR103-105,107-119
445. O6N2_HUMAN (Q8NGY6) OR6N2 Olfactory II fam 6 / MOR103-105,107-119
446. O6P1_HUMAN (Q8NGX9) OR6P1 Olfactory II fam 6 / MOR103-105,107-119
447. O6Q1_HUMAN (Q8NGQ2) OR6Q1 Olfactory II fam 6 / MOR103-105,107-119
448. O6S1_HUMAN (Q8NH40) OR6S1 Olfactory II fam 6 / MOR103-105,107-119
449. O6T1_HUMAN (Q8NGN1) OR6T1 Olfactory II fam 6 / MOR103-105,107-119
450. O6V1_HUMAN (Q8N148) OR6V1 Olfactory II fam 6 / MOR103-105,107-119
451. O6W1_HUMAN (Q96RR8) OR6W1 fragment
452.O6X1_HUMAN (Q8NH79) OR6X1 Olfactory II fam 6 / MOR103-105,107-119
453. O6Y1_HUMAN (Q8NGX8) OR6Y1 Olfactory II fam 6 / MOR103-105,107-119
454. O75171 (O75171) KIAA0686 Estimated / Unclassified Class B GPCRs
455. O75205 (O75205) A-69G12.1 OR GPRC5B Orphan GPRC5
456. O75228 (O75228) TBXA2R Thromboxane
457. O75303 (O75303) CCR-5 fragment
458. O75307 (O75307) CRAM-A CC Chemokine and others
459. O75824 (O75824) CCKBR Fragment
460. O75963 (O75963) none Estimated / Unclassified Class A GPCRs
461. O7A2_HUMAN (Q8NGA2) OR7A2 OR OR7A7 Olfactory II fam 7 / MOR139-155
462. O7A5_HUMAN (Q15622) OR7A5 Olfactory II fam 7 / MOR139-155
463. O7AA_HUMAN (O76100) OR7A10 Olfactory II fam 7 / MOR139-155
464. O7AH_HUMAN (O14581) OR7A17 Olfactory II fam 7 / MOR139-155
465. O7C1_HUMAN (O76099) OR7C1 Olfactory II fam 7 / MOR139-155
466. O7C2_HUMAN (O60412) OR7C2 OR OR7C3 Olfactory II fam 7 / MOR139-155
467. O7D2_HUMAN (Q96RA2) OR7D2 Olfactory II fam 7 / MOR139-155
468. O7G1_HUMAN (Q8NGA0) OR7G1 Olfactory II fam 7 / MOR139-155
469. O7G2_HUMAN (Q8NG99) OR7G2 Olfactory II fam 7 / MOR139-155
470. O7G3_HUMAN (Q8NG95) OR7G3 Olfactory II fam 7 / MOR139-155
471. O8A1_HUMAN (Q8NGG7) OR8A1 Olfactory II fam 8 / MOR161-171
472. O8B2_HUMAN (Q96RD0) OR8B2 Olfactory II fam 8 / MOR161-171
473. O8B3_HUMAN (Q8NGG8) OR8B3 Olfactory II fam 8 / MOR161-171
474. O8B4_HUMAN (Q96RC9) OR8B4 Olfactory II fam 8 / MOR161-171
475. O8B8_HUMAN (Q15620) OR8B8 Olfactory II fam 8 / MOR161-171
476. O8BC_HUMAN (Q8NGG6) OR8B12 Olfactory II fam 8 / MOR161-171
477. O8D1_HUMAN (Q8WZ84) OR8D1 Olfactory II fam 8 / MOR161-171
478. O8D2_HUMAN (Q9GZM6) OR8D2 Olfactory II fam 8 / MOR161-171
479. O8D4_HUMAN (Q8NGM9) OR8D4 Olfactory II fam 8 / MOR161-171
480. O8G1_HUMAN (Q15617) OR8G1 fragment
481. O8G2_HUMAN (Q15614) OR8G2 fragment
482. O8H1_HUMAN (Q8NGG4) OR8H1 Olfactory II fam 5 / MOR172-224,249,254
483. O8H2_HUMAN (Q8N162) OR8H2 Olfactory II fam 5 / MOR172-224,249,254
484. O8H3_HUMAN (Q8N146) OR8H3 Olfactory II fam 5 / MOR172-224,249,254
485. O8I2_HUMAN (Q8N0Y5) OR8I2 Olfactory II fam 5 / MOR172-224,249,254
486. O8J1_HUMAN (Q8NGP2) OR8J1 Olfactory II fam 5 / MOR172-224,249,254
487. O8J3_HUMAN (Q8NGG0) OR8J3 Olfactory II fam 5 / MOR172-224,249,254
488. O8K1_HUMAN (Q8NGG5) OR8K1 Olfactory II fam 5 / MOR172-224,249,254
489. O8K3_HUMAN (Q8NH51) OR8K3 Olfactory II fam 5 / MOR172-224,249,254
490. O8K5_HUMAN (Q8NH50) OR8K5 Olfactory II fam 5 / MOR172-224,249,254
491. O94858 (O94858) KIAA0758 Estimated / Unclassified Class B GPCRs
492. O94867 (O94867) KIAA0768 fragment
493.O94882 (O94882) KIAA0786 fragment
494. O94910 (O94910) KIAA0821 Latrophilin Type 1
495. O95220 (O95220) OR5D3 fragment
496. O95357 (O95357) RAIG1 Orphan GPRC5
497. O95490 (O95490) LPHH1 OR LEC1 Latrophilin Type 2
498. O95499 (O95499) OLFR89 Olfactory II fam 2 / MOR256-262,270-285
499. O95800 (O95800) none Estimated / Unclassified Class A GPCRs
500. O95950 (O95950) none fragment
501. O95966 (O95966) none Estimated / Unclassified Class B GPCRs
502. O95977 (O95977) EDG6 Lysosphingolipid & LPA (EDG)
503. O9A2_HUMAN (Q8NGT5) OR9A2 Olfactory II fam 9 / MOR120
504. O9A4_HUMAN (Q8NGU2) OR9A4 Olfactory II fam 9 / MOR120
505. O9G1_HUMAN (Q8NH87) OR9G1 Olfactory II fam 9 / MOR120
506. O9G4_HUMAN (Q8NGQ1) OR9G4 Olfactory II fam 5 / MOR172-224,249,254
507. O9G5_HUMAN (Q8NGQ0) OR9G5 Olfactory II fam 9 / MOR120
508. O9I1_HUMAN (Q8NGQ6) OR9I1 Olfactory II fam 5 / MOR172-224,249,254
509. O9K2_HUMAN (Q8NGE7) OR9K2 Olfactory II fam 5 / MOR172-224,249,254
510. O9Q1_HUMAN (Q8NGQ5) OR9Q1 Olfactory II fam 9 / MOR120
511. OA1_HUMAN (P51810) OA1 Ocular albinism proteins
512. OAA1_HUMAN (O95223) OR10A1 fragment
513. OAA3_HUMAN (P58181) OR10A3 Olfactory II fam 10 / MOR263-269
514. OAA4_HUMAN (Q9H209) OR10A4 Olfactory II fam 10 / MOR263-269
515. OAA5_HUMAN (Q9H207) OR10A5 Olfactory II fam 10 / MOR263-269
516. OAA6_HUMAN (Q8NH74) OR10A6 Olfactory II fam 10 / MOR263-269
517. OAA7_HUMAN (Q8NGE5) OR10A7 Olfactory II fam 10 / MOR263-269
518. OAC1_HUMAN (Q96KK4) OR10C1 OR OR10C2 Olfactory II fam 10 / MOR263-269
519. OAD1_HUMAN (Q8NGE0) OR10AD1 Olfactory II fam 10 / MOR263-269
520. OAD4_HUMAN (Q8NGN7) OR10D4 Olfactory II fam 5 / MOR172-224,249,254
521. OAG1_HUMAN (Q8NH19) OR10AG1 Olfactory II fam 10 / MOR263-269
522. OAG2_HUMAN (Q8NGC3) OR10G2 Olfactory II fam 5 / MOR172-224,249,254
523. OAG3_HUMAN (Q8NGC4) OR10G3 Olfactory II fam 5 / MOR172-224,249,254
524. OAG4_HUMAN (Q8NGN3) OR10G4 Olfactory II fam 5 / MOR172-224,249,254
525. OAG7_HUMAN (Q8NGN6) OR10G7 Olfactory II fam 5 / MOR172-224,249,254
526. OAG8_HUMAN (Q8NGN5) OR10G8 Olfactory II fam 5 / MOR172-224,249,254
527. OAG9_HUMAN (Q8NGN4) OR10G9 Olfactory II fam 5 / MOR172-224,249,254
528. OAH1_HUMAN (Q9Y4A9) OR10H1 Olfactory II fam 10 / MOR263-269
529. OAH2_HUMAN (O60403) OR10H2 Olfactory II fam 10 / MOR263-269
530. OAH3_HUMAN (O60404) OR10H3 Olfactory II fam 10 / MOR263-269
531.OAH4_HUMAN (Q8NGA5) OR10H4 Olfactory II fam 10 / MOR263-269
532. OAH5_HUMAN (Q8NGA6) OR10H5 Olfactory II fam 10 / MOR263-269
533. OAJ1_HUMAN (P30954) OR10J1 Olfactory II fam 10 / MOR263-269
534.OAJ5_HUMAN (Q8NHC4) OR10J5 Olfactory II fam 10 / MOR263-269
535. OAJ6_HUMAN (Q8NGY7) OR10J6 Olfactory II fam 10 / MOR263-269
536. OAK1_HUMAN (Q8NGX5) OR10K1 Olfactory II fam 10 / MOR263-269
537. OAQ1_HUMAN (Q8NGQ4) OR10Q1 Olfactory II fam 10 / MOR263-269
538. OAR2_HUMAN (Q8NGX6) OR10R2 Olfactory II fam 10 / MOR263-269
539. OASI_HUMAN (Q8NGN2) OR10S1 Olfactory II fam 5 / MOR172-224,249,254
540. OAT2_HUMAN (Q8NGX3) OR10T2 Olfactory II fam 10 / MOR263-269
541. OAV1_HUMAN (Q8NGI7) OR10V1 Olfactory II fam 10 / MOR263-269
542. OAZ1_HUMAN (Q8NGY1) OR10Z1 Olfactory II fam 10 / MOR263-269
543. OBA1_HUMAN (Q9GZK7) OR11A1 OR OR11A2 Olfactory II fam 11 / MOR106,121-122
544. OBG2_HUMAN (Q8NGC1) OR11G2 Olfactory II fam 11 / MOR106,121-122
545.OBH1_HUMAN (Q8NG94) OR11H1 Olfactory II fam 11 / MOR106,121-122
546.OBH4_HUMAN (Q8NGC9) OR11H4 Olfactory II fam 11 / MOR106,121-122
547.OBH6_HUMAN (Q8NGC7) OR11H6 Olfactory II fam 11 / MOR106,121-122
548. OBL1_HUMAN (Q8NGX0) OR11L1 Olfactory II fam 6 / MOR103-105,107-119
549. OCD2_HUMAN (P58182) OR12D2 Olfactory II fam 12 / MOR250
550. OCD3_HUMAN (Q9UGF7) OR12D3 Olfactory II fam 12 / MOR250
551. ODA1_HUMAN (Q8NGR1) OR13A1 Olfactory II fam 13 / MOR253
552. ODC2_HUMAN (Q8NGS9) OR13C2 Olfactory II fam 2 / MOR256-262,270-285
553. ODC3_HUMAN (Q8NGS6) OR13C3 Olfactory II fam 2 / MOR256-262,270-285
554. ODC4_HUMAN (Q8NGS5) OR13C4 Olfactory II fam 2 / MOR256-262,270-285
555. ODC5_HUMAN (Q8NGS8) OR13C5 Olfactory II fam 2 / MOR256-262,270-285
556. ODC8_HUMAN (Q8NGS7) OR13C8 Olfactory II fam 2 / MOR256-262,270-285
557. ODC9_HUMAN (Q8NGT0) OR13C9 Olfactory II fam 2 / MOR256-262,270-285
558. ODD1_HUMAN (Q8NGV5) OR13D1 Olfactory II fam 2 / MOR256-262,270-285
559. ODF1_HUMAN (Q8NGS4) OR13F1 Olfactory II fam 2 / MOR256-262,270-285
560. ODG1_HUMAN (Q8NGZ3) OR13G1 Olfactory II fam 13 / MOR253
561. ODH1_HUMAN (Q8NG92) OR13H1 Olfactory II fam 13 / MOR253
562.ODJ1_HUMAN (Q8NGT2) OR13J1 Olfactory II fam 2 / MOR256-262,270-285
563. OK4E_HUMAN (Q8NGD5) OR4K14 Olfactory II fam 4 / MOR225-248
564. OPN3_HUMAN (Q9H1Y3) OPN3 OR ECPN Rhodopsin Other
565. OPN4_HUMAN (Q9UHM6) OPN4 OR MOP Rhodopsin etc.
566. OPRD_HUMAN (P41143) OPRD1 OR OPRD Opioid Type D
567. OPRK_HUMAN (P41145) OPRK1 OR OPRK Opioid Type K
568. OPRM_HUMAN (P35372) OPRM1 OR MOR1 Opioid Type M
569. OPRX_HUMAN (P41146) OPRL1 OR ORL1 OR OOR opioid type X
570. OPSB_HUMAN (P03999) OPN1SW OR BCP Rhodopsin Vertebrate Type 3
571. OPSG_HUMAN (P04001) OPN1MW OR GCP Rhodopsin Vertebrate Type 2
572. OPSX_HUMAN (O14718) RRH Rhodopsin and others
573. OX1R_HUMAN (O43613) HCRTR1 Orexin
574. OX2R_HUMAN (O43614) HCRTR2 Orexin
575. OXA2_HUMAN (Q8NGJ7) OR51A2 Olfactory I fam 51-52 / MOR1-42
576. OXA4_HUMAN (Q8NGJ6) OR51A4 Olfactory I fam 51-52 / MOR1-42
577. OXA7_HUMAN (Q8NH64) OR51A7 Olfactory I fam 51-52 / MOR1-42
578. OXB2_HUMAN (Q9Y5P1) OR51B2 Olfactory I fam 51-52 / MOR1-42
579. OXB4_HUMAN (Q9Y5P0) OR51B4 Olfactory I fam 51-52 / MOR1-42
580. OXE1_HUMAN (Q8TCB6) OR51E1 Olfactory I fam 51-52 / MOR1-42
581. OXE2_HUMAN (Q9H255) OR51E2 OR PSGR Olfactory I fam 51-52 / MOR1-42
582. OXF2_HUMAN (Q8NH61) OR51F2 Olfactory I fam 51-52 / MOR1-42
583. OXG1_HUMAN (Q8NGK1) OR51G1 Olfactory I fam 51-52 / MOR1-42
584. OXG2_HUMAN (Q8NGK0) OR51G2 Olfactory I fam 51-52 / MOR1-42
585. OXH1_HUMAN (Q8NH63) OR51H1 Olfactory I fam 51-52 / MOR1-42
586. OXI1_HUMAN (Q9H343) OR51I1 Olfactory I fam 51-52 / MOR1-42
587. OXI2_HUMAN (Q9H344) OR51I2 Olfactory I fam 51-52 / MOR1-42
588. OXL1_HUMAN (Q8NGJ5) OR51L1 Olfactory I fam 51-52 / MOR1-42
589. OXM1_HUMAN (Q9H341) OR51M1 Olfactory I fam 51-52 / MOR1-42
590. OXQ1_HUMAN (Q8NH59) OR51Q1 Olfactory I fam 51-52 / MOR1-42
591. OXS1_HUMAN (Q8NGJ8) OR51S1 Olfactory I fam 51-52 / MOR1-42
592. OXT1_HUMAN (Q8NGJ9) OR51T1 Olfactory I fam 51-52 / MOR1-42
593. OXYR_HUMAN (P30559) OXTR Oxytocin
594. OYA1_HUMAN (Q9UKL2) OR52A1 Olfactory I fam 51-52 / MOR1-42
595. OYB2_HUMAN (Q96RD2) OR52B2 Olfactory I fam 51-52 / MOR1-42
596. OYB4_HUMAN (Q8NGK2) OR52B4 Olfactory I fam 51-52 / MOR1-42
597. OYB6_HUMAN (Q8NGF0) OR52B6 Olfactory I fam 51-52 / MOR1-42
598. OYD1_HUMAN (Q9H346) OR52D1 Olfactory I fam 51-52 / MOR1-42
599. OYE1_HUMAN (Q8NGJ3) OR52E1 Olfactory I fam 51-52 / MOR1-42
600. OYE2_HUMAN (Q8NGJ4) OR52E2 Olfactory I fam 51-52 / MOR1-42
601. OYE4_HUMAN (Q8NGH9) OR52E4 Olfactory I fam 51-52 / MOR1-42
602. OYE5_HUMAN (Q8NH55) OR52E5 Olfactory I fam 51-52 / MOR1-42
603. OYE6_HUMAN (Q96RD3) OR52E6 Olfactory I fam 51-52 / MOR1-42
604. OYH1_HUMAN (Q8NGJ2) OR52H1 Olfactory I fam 51-52 / MOR1-42
605. OYI1_HUMAN (Q8NGK6) OR52I1 Olfactory I fam 51-52 / MOR1-42
606. OYI2_HUMAN (Q8NH67) OR52I2 Olfactory I fam 51-52 / MOR1-42
607. OYJ3_HUMAN (Q8NH60) OR52J3 Olfactory I fam 51-52 / MOR1-42
608. OYK1_HUMAN (Q8NGK4) OR52K1 Olfactory I fam 51-52 / MOR1-42
609. OYK2_HUMAN (Q8NGK3) OR52K2 Olfactory I fam 51-52 / MOR1-42
610. OYL1_HUMAN (Q8NGH7) OR52L1 Olfactory I fam 51-52 / MOR1-42
611. OYL2_HUMAN (Q8NGH6) OR52L2 Olfactory I fam 51-52 / MOR1-42
612. OYN1_HUMAN (Q8NH53) OR52N1 Olfactory I fam 51-52 / MOR1-42
613. OYN2_HUMAN (Q8NGI0) OR52N2 Olfactory I fam 51-52 / MOR1-42
614. OYN4_HUMAN (Q8NGI2) OR52N4 Olfactory I fam 51-52 / MOR1-42
615. OYN5_HUMAN (Q8NH56) OR52N5 Olfactory I fam 51-52 / MOR1-42
616. OYP1_HUMAN (Q8NH57) OR52P1 Olfactory I fam 51-52 / MOR1-42
617. OYR1_HUMAN (Q8NGF1) OR52R1 Olfactory I fam 51-52 / MOR1-42
618. OZA1_HUMAN (Q8NGH5) OR56A1 Olfactory I fam 51-52 / MOR1-42
619. OZA4_HUMAN (Q8NGH8) OR56A4 Olfactory I fam 51-52 / MOR1-42
620. OZB2_HUMAN (Q8NGI1) OR56B2 Olfactory I fam 51-52 / MOR1-42
621. OZB4_HUMAN (Q8NH76) OR56B4 Olfactory I fam 51-52 / MOR1-42
622. P2Y2_HUMAN (P41231) P2RY2 OR P2RU1 Purine Receptor Type U
623. P2Y4_HUMAN (P51582) P2RY4 OR NRU Purinoceptor type Y adenine / uridine
624. P2Y5_HUMAN (P43657) P2RY5 Purine Receptor etc.
625. P2Y6_HUMAN (Q15077) P2RY6 Purine Receptor Type Y Adenine / Uridine
626. P2Y9_HUMAN (Q99677) GPR23 OR P2RY9 Purine Receptor etc.
627. P2YA_HUMAN (O00398) P2RY10 Purine receptor etc.
628. P2YB_HUMAN (Q96G91) P2RY11 Purine Receptor etc.
629. P2YC_HUMAN (Q9H244) P2RY12 OR HORK3 Platelet ADP & KI01 Receptors
630. P2YR_HUMAN (P47900) P2RY1 Purine receptor type Y adenine / uridine
631. P2YX_HUMAN (Q15391) GPR105 OR KIAA0001 Platelet ADP & KI01 Receptors
632. P78470 (P78470) none fragment
633. P78471 (P78471) none fragment
634. PACR_HUMAN (P41586) ADCYAP1R1 PACAP
635. PAFR_HUMAN (P25105) PTAFR OR PAFR Platelet activating factor
636.PAR1_HUMAN (P25116) F2R OR PAR1 OR TR OR CF2R Thrombin
637. PAR2_HUMAN (P55085) F2RL1 OR PAR2 OR GPR11 Proteinase-activated
638. PAR3_HUMAN (O00254) F2RL2 OR PAR3 proteinase activation
639. PAR4_HUMAN (Q96RI0) F2RL3 OR PAR4 proteinase activation
640. PD2R_HUMAN (Q13258) PTGDR Prostaglandin E2 / D2 Subtype EP2
641. PE21_HUMAN (P34995) PTGER1 Prostaglandin E2 Subtype EP1
642. PE22_HUMAN (P43116) PTGER2 Prostaglandin E2 / D2 Subtype EP2
643. PE23_HUMAN (P43115) PTGER3 Prostaglandin E2 Subtype EP3
644. PE24_HUMAN (P35408) PTGER4 OR PTGER2 Prostaglandin E2 Subtype EP4
645. PF2R_HUMAN (P43088) PTGFR Prostaglandin F2-alpha
646. PI2R_HUMAN (P43119) PTGIR OR PRIPR Prostacyclin
647. PKR1_HUMAN (Q8TCW9) GPR73 OR PKR1 Prokineticin receptors
648. PKR2_HUMAN (Q8NFJ6) GPR73L1 OR PKR2 Prokineticin Receptors
649. PRK2_HUMAN (Q9HC23) PROK2 OR BV8 fragment
650. PSYR_HUMAN (Q8IYL9) GPR65 OR TDAG8 Estimated / Unclassified Class A GPCRs
651. PTR2_HUMAN (P49190) PTHR2 Parathyroid hormone
652. PTRR_HUMAN (Q03431) PTHR1 OR PTHR Parathyroid hormone
653.Q13027 (Q13027) none fragment
654. Q13167 (Q13167) DRD3 Dopamine Vertebrate Type 3
655. Q13675 (Q13675) none α-adrenergic receptor type 1
656. Q13729 (Q13729) none α-adrenergic receptor type 1
657. Q14439 (Q14439) none Estimated / Unclassified Class A GPCRs
658. Q14968 (Q14968) none Estimated / Unclassified Class A GPCRs
659. Q15613 (Q15613) TPCR110 fragment
660. Q15616 (Q15616) TPCR24 fragment
661. Q15618 (Q15618) TPCR26 fragment
662.Q16144 (Q16144) none CCK Type B
663.Q16292 (Q16292) THROMBIN RECEPTOR fragment
664. Q16303 (Q16303) none fragment
665. Q16432 (Q16432) none fragment
666. Q16433 (Q16433) none fragment
667. Q16503 (Q16503) none fragment
668. Q16538 (Q16538) A-2 Estimated / Unclassified Class A GPCRs
669. Q7Z3S0 (Q7Z3S0) DKFZP686B1664 Purine Receptor etc.
670. Q7Z3S6 (Q7Z3S6) DKFZP686P1665 Purine receptor etc.
671. Q7Z3T1 (Q7Z3T1) DKFZP686N09198 Olfactory II fam 2 / MOR256-262,270-285
672. Q7Z3W3 (Q7Z3W3) DKFZP686N1782 Proteinase activation
673. Q7Z582 (Q7Z582) GPR50 fragment
674. Q7Z5J9 (Q7Z5J9) GPRC5D Orphan GPRC5
675. Q7Z5R9 (Q7Z5R9) none Histamine Type 2
676. Q7Z601 (Q7Z601) GPR142 Estimated / Unclassified Class A GPCRs
677. Q7Z603 (Q7Z603) GPR136 Estimated / Unclassified Class A GPCRs
678. Q7Z604 (Q7Z604) GPR135 Estimated / Unclassified Class A GPCRs
679. Q7Z605 (Q7Z605) GPR120 Estimated / Unclassified Class A GPCRs
680. Q7Z710 (Q7Z710) GPR9 CXC Chemokine Type 3
681. Q7Z7I1 (Q7Z7I1) CCBP2 CC Chemokine Type X
682. Q7Z7M1 (Q7Z7M1) GPR144 Estimated / Unclassified Class B GPCRs
683. Q7Z7M2 (Q7Z7M2) GPR133 Estimated / Unclassified Class B GPCRs
684. Q7Z7Q5 (Q7Z7Q5) DRD4 fragment
685. Q86SE3 (Q86SE3) none fragment
686. Q86SF1 (Q86SF1) none fragment
687. Q86SF3 (Q86SF3) none fragment
688. Q86SF4 (Q86SF4) none fragment
689. Q86SG0 (Q86SG0) none fragment
690. Q86SG8 (Q86SG8) none fragment
691. Q86SG9 (Q86SG9) none fragment
692. Q86SH1 (Q86SH1) none fragment
693. Q86SH3 (Q86SH3) none fragment
694. Q86SI3 (Q86SI3) none fragment
695. Q86SI5 (Q86SI5) none fragment
696. Q86SI8 (Q86SI8) none fragment
697. Q86SJ4 (Q86SJ4) none fragment
698. Q86SL3 (Q86SL3) none fragment
699. Q86SL4 (Q86SL4) none fragment
700. Q86SL7 (Q86SL7) none fragment
701. Q86SL9 (Q86SL9) none fragment
702. Q86SM0 (Q86SM0) none fragment
703. Q86SM1 (Q86SM1) none fragment
704. Q86SM3 (Q86SM3) none fragment
705. Q86SM4 (Q86SM4) none fragment
706. Q86SM7 (Q86SM7) none fragment
707. Q86SN8 (Q86SN8) none fragment
708. Q86SN9 (Q86SN9) none fragment
709. Q86SP1 (Q86SP1) none fragment
710. Q86SP4 (Q86SP4) none fragment
711. Q86SP7 (Q86SP7) none fragment
712. Q86SQ2 (Q86SQ2) GPR128 fragment
713. Q86SQ3 (Q86SQ3) GPR127 Fragment
714. Q86SQ4 (Q86SQ4) GPR126 Estimated / Unclassified Class B GPCRs
715. Q86SQ5 (Q86SQ5) GPR125 fragment
716. Q86SQ6 (Q86SQ6) GPR123 Estimated / Unclassified Class B GPCRs
717. Q86TH8 (Q86TH8) none α-adrenergic receptor type 2
718. Q86U87 (Q86U87) none Estimated / Unclassified Class A GPCRs
719. Q86UG6 (Q86UG6) none fragment
720. Q86UG7 (Q86UG7) none fragment
721. Q86UG8 (Q86UG8) none fragment
722. Q86UG9 (Q86UG9) none fragment
723. Q86UH0 (Q86UH0) none fragment
724. Q86UN1 (Q86UN1) HTR5A Fragment
725. Q86UN7 (Q86UN7) CASR fragment
726. Q86UZ8 (Q86UZ8) FZD2 fragment
727. Q86V16 (Q86V16) none Purine receptor etc.
728. Q86V80 (Q86V80) none Opioid Type M
729. Q86VA9 (Q86VA9) CCR1 CC Chemokine Type 1
730. Q86VP7 (Q86VP7) none GPR37 / Endothelin B-like
731. Q86VZ1 (Q86VZ1) none Estimated / Unclassified Class A GPCRs
732. Q86W60 (Q86W60) none GABA-B Subtype 1
733. Q86WP7 (Q86WP7) none Estimated / Unclassified Class A GPCRs
734. Q86XI5 (Q86XI5) none fragment
735. Q86Y34 (Q86Y34) GPR97 Estimated / Unclassified Class B GPCRs
736. Q86YF2 (Q86YF2) none fragment
737. Q86YG3 (Q86YG3) none fragment
738. Q86YG6 (Q86YG6) none Metabotropic Glutamic Acid Group II
739. Q86YG9 (Q86YG9) none fragment
740. Q86YW1 (Q86YW1) MC1R Melanocyte Stimulating Hormone
741. Q8IU63 (Q8IU63) 6M1-16 Fragment
742. Q8IUZ1 (Q8IUZ1) none Interleukin-8 Type B
743. Q8IV06 (Q8IV06) none Estimated / Unclassified Class A GPCRs
744. Q8IV17 (Q8IV17) none fragment
745. Q8IV19 (Q8IV19) none Cysteinyl leukotrienes
746. Q8IVW0 (Q8IVW0) none Acetylcholine Vertebrate Type 5
747. Q8IW08 (Q8IW08) none GABA-B Subtype 1
748. Q8IWK6 (Q8IWK6) none fragment
749. Q8IWP3 (Q8IWP3) none Opioid Type K
750. Q8IWT2 (Q8IWT2) none Estimated / Unclassified Class B GPCRs
751. Q8IWW3 (Q8IWW3) OPRM Opioid Type M
752. Q8IWW4 (Q8IWW4) OPRM opioid type M
753. Q8IX08 (Q8IX08) OPN3 fragment
754. Q8IXA4 (Q8IXA4) VIGR Estimated / Unclassified Class B GPCRs
755. Q8IXB0 (Q8IXB0) none Opioid Type X
756. Q8IXD8 (Q8IXD8) none Estimated / Unclassified Class B GPCRs
757. Q8IXD9 (Q8IXD9) none fragment
758. Q8IXE0 (Q8IXE0) none fragment
759. Q8IXE2 (Q8IXE2) none Estimated / Unclassified Class A GPCRs
760. Q8IXE3 (Q8IXE3) none Estimated / Unclassified Class B GPCRs
761. Q8IXE4 (Q8IXE4) none Estimated / Unclassified Class B GPCRs
762. Q8IXE5 (Q8IXE5) none Estimated / Unclassified Class A GPCRs
763. Q8IXE6 (Q8IXE6) none Estimated / Unclassified Class B GPCRs
764. Q8IXE7 (Q8IXE7) none Olfactory II fam 5 / MOR172-224,249,254
765. Q8IXH9 (Q8IXH9) HTR4B Serotonin Vertebrate Type 4
766. Q8IY58 (Q8IY58) none Neurotensin
767. Q8IYT7 (Q8IYT7) none Platelet ADP & KI01 receptors
768. Q8IZ08 (Q8IZ08) none Estimated / Unclassified Class A GPCRs
769. Q8IZ14 (Q8IZ14) none Estimated / Unclassified Class B GPCRs
770. Q8IZE4 (Q8IZE4) none Estimated / Unclassified Class B GPCRs
771. Q8IZE5 (Q8IZE5) none Estimated / Unclassified Class B GPCRs
772. Q8IZE6 (Q8IZE6) none Estimated / Unclassified Class B GPCRs
773. Q8IZE7 (Q8IZE7) none Estimated / Unclassified Class B GPCRs
774. Q8IZF1 (Q8IZF1) GPR97 Estimated / Unclassified Class B GPCRs
775. Q8IZF2 (Q8IZF2) GPR116 Estimated / Unclassified Class B GPCRs
776. Q8IZF3 (Q8IZF3) GPR115 Estimated / Unclassified Class B GPCRs
777. Q8IZF4 (Q8IZF4) GPR114 Estimated / Unclassified Class B GPCRs
778. Q8IZF5 (Q8IZF5) GPR113 Estimated / Unclassified Class B GPCRs
779. Q8IZF6 (Q8IZF6) GPR112 Estimated / Unclassified Class B GPCRs
780. Q8IZF7 (Q8IZF7) GPR111 Estimated / Unclassified Class B GPCRs
781. Q8IZF8 (Q8IZF8) GPR110 Estimated / Unclassified Class B GPCRs
782. Q8IZP3 (Q8IZP3) GPR64 Estimated / Unclassified Class B GPCRs
783. Q8IZP4 (Q8IZP4) GPR64 Estimated / Unclassified Class B GPCRs
784. Q8IZP9 (Q8IZP9) none Estimated / Unclassified Class B GPCRs
785. Q8N0W0 (Q8N0W0) none fragment
786. Q8N0W1 (Q8N0W1) none fragment
787. Q8N0X1 (Q8N0X1) none fragment
788. Q8N0Y1 (Q8N0Y1) none Olfactory II fam 8 / MOR161-171
789. Q8N0Z0 (Q8N0Z0) none fragment
790. Q8N164 (Q8N164) none fragment
791. Q8N261 (Q8N261) none fragment
792. Q8N2R3 (Q8N2R3) none fragment
793. Q8N429 (Q8N429) none fragment
794. Q8N537 (Q8N537) none LGR-like (hormone receptors)
795. Q8N580 (Q8N580) none Estimated / Unclassified Class A GPCRs
796. Q8N5S7 (Q8N5S7) none GPR
797. Q8N5V1 (Q8N5V1) none fragment
798. Q8N639 (Q8N639) none Estimated / Unclassified Class B GPCRs
799. Q8N6T6 (Q8N6T6) none Interleukin-8 Type A
800. Q8N6U8 (Q8N6U8) none Estimated / Unclassified Class A GPCRs
801. Q8N7J6 (Q8N7J6) none Estimated / Unclassified Class A GPCRs
802. Q8NBZ8 (Q8NBZ8) none Orphan GPRC5
803. Q8NC14 (Q8NC14) none fragment
804. Q8NCH4 (Q8NCH4) none fragment
805. Q8NE10 (Q8NE10) none RDC1
806. Q8NE20 (Q8NE20) none Neuromedin U
807. Q8NEI9 (Q8NEI9) none fragment
808. Q8NEN2 (Q8NEN2) none SREB
809. Q8NFJ5 (Q8NFJ5) none Orphan GPRC5
810. Q8NFN8 (Q8NFN8) GABABL GABA-B like (GABA-B like)
811. Q8NFS2 (Q8NFS2) GRM7 Metabotropic Glutamate Group III
812. Q8NFS3 (Q8NFS3) GRM7 Metabotropic Glutamate Group III
813. Q8NFS4 (Q8NFS4) GRM7 Metabotropic Glutamate Group III
814. Q8NG71 (Q8NG71) none Corticotropin releasing factor
815.Q8NG72 (Q8NG72) none EMR1
816. Q8NG73 (Q8NG73) none fragment
817. Q8NG75 (Q8NG75) none Olfactory II fam 5 / MOR172-224,249,254
818. Q8NG76 (Q8NG76) none Olfactory II fam 2 / MOR256-262,270-285
819.Q8NG77 (Q8NG77) none Olfactory II fam 2 / MOR256-262,270-285
820. Q8NG78 (Q8NG78) none Olfactory II fam 8 / MOR161-171
821. Q8NG79 (Q8NG79) none Olfactory unclassified class II
822. Q8NG82 (Q8NG82) none Olfactory II fam 2 / MOR256-262,270-285
823. Q8NG85 (Q8NG85) none Olfactory II fam 2 / MOR256-262,270-285
824. Q8NG86 (Q8NG86) none Olfactory II fam 2 / MOR256-262,270-285
825. Q8NG87 (Q8NG87) none fragment
826.Q8NG88 (Q8NG88) none Olfactory II fam 8 / MOR161-171
827. Q8NG89 (Q8NG89) none fragment
828. Q8NG90 (Q8NG90) none fragment
829. Q8NG91 (Q8NG91) none fragment
830. Q8NG96 (Q8NG96) none EMR1
831.Q8NG98 (Q8NG98) none Olfactory II fam 7 / MOR139-155
832. Q8NGA3 (Q8NGA3) none fragment
833. Q8NGA4 (Q8NGA4) none Chemokine receptor-like 1
834. Q8NGA7 (Q8NGA7) none EMR1
835. Q8NGA9 (Q8NGA9) none Estimated / Unclassified Class B GPCRs
836. Q8NGB0 (Q8NGB0) none Estimated / Unclassified Class A GPCRs
837. Q8NGB2 (Q8NGB2) none Olfactory II fam 4 / MOR225-248
838. Q8NGB3 (Q8NGB3) none Estimated / Unclassified Class B GPCRs
839. Q8NGB5 (Q8NGB5) none fragment
840. Q8NGC8 (Q8NGC8) none fragment
841.Q8NGD6 (Q8NGD6) none Olfactory II fam 4 / MOR225-248
842. Q8NGD7 (Q8NGD7) none fragment
843. Q8NGD8 (Q8NGD8) none fragment
844. Q8NGD9 (Q8NGD9) none Olfactory II fam 9 / MOR120
845. Q8NGE2 (Q8NGE2) none Olfactory II fam 6 / MOR103-105,107-119
846. Q8NGE3 (Q8NGE3) none Olfactory II fam 10 / MOR263-269
847. Q8NGE4 (Q8NGE4) none Estimated / Unclassified Class A GPCRs
848. Q8NGE6 (Q8NGE6) none fragment
849.Q8NGE9 (Q8NGE9) none Olfactory II fam 5 / MOR172-224,249,254
850. Q8NGF2 (Q8NGF2) none fragment
851. Q8NGF3 (Q8NGF3) none Olfactory I fam 51-52 / MOR1-42
852. Q8NGF6 (Q8NGF6) none Olfactory II fam 10 / MOR263-269
853. Q8NGG1 (Q8NGG1) none fragment
854. Q8NGG9 (Q8NGG9) none Olfactory II fam 8 / MOR161-171
855. Q8NGH0 (Q8NGH0) none Olfactory II fam 8 / MOR161-171
856. Q8NGH1 (Q8NGH1) none Olfactory II fam 8 / MOR161-171
857. Q8NGH2 (Q8NGH2) none Olfactory II fam 8 / MOR161-171
858. Q8NGH4 (Q8NGH4) none Olfactory II fam 2 / MOR256-262,270-285
859. Q8NGI3 (Q8NGI3) none Olfactory I fam 51-52 / MOR1-42
860. Q8NGI4 (Q8NGI4) none Olfactory II fam 4 / MOR225-248
861. Q8NGI5 (Q8NGI5) none fragment
862. Q8NGI6 (Q8NGI6) none Olfactory II fam 4 / MOR225-248
863. Q8NGK5 (Q8NGK5) none Olfactory I fam 51-52 / MOR1-42
864. Q8NGK7 (Q8NGK7) none Mas-related receptors (MRGs)
865. Q8NGK8 (Q8NGK8) none fragment
866. Q8NGL5 (Q8NGL5) none Olfactory II fam 5 / MOR172-224,249,254
867. Q8NGL8 (Q8NGL8) none Olfactory II fam 4 / MOR225-248
868. Q8NGM0 (Q8NGM0) none Olfactory II fam 4 / MOR225-248
869. Q8NGM3 (Q8NGM3) none fragment
870. Q8NGM4 (Q8NGM4) none Olfactory II fam 10 / MOR263-269
871. Q8NGM5 (Q8NGM5) none Dopamine Vertebrate Type 4
872. Q8NGM6 (Q8NGM6) none fragment
873. Q8NGM7 (Q8NGM7) none Olfactory I fam 51-52 / MOR1-42
874. Q8NGN9 (Q8NGN9) none Olfactory II fam 4 / MOR225-248
875. Q8NGP1 (Q8NGP1) none fragment
876. Q8NGP5 (Q8NGP5) none fragment
877. Q8NGP7 (Q8NGP7) none fragment
878. Q8NGQ7 (Q8NGQ7) none fragment
879. Q8NGQ8 (Q8NGQ8) none Substance K (NK2)
880. Q8NGQ9 (Q8NGQ9) none Rhodopsin etc.
881. Q8NGT3 (Q8NGT3) none fragment
882. Q8NGT4 (Q8NGT4) none fragment
883. Q8NGT6 (Q8NGT6) none Olfactory II fam 2 / MOR256-262,270-285
884. Q8NGT8 (Q8NGT8) none Olfactory II fam 2 / MOR256-262,270-285
885. Q8NGU0 (Q8NGU0) none fragment
886. Q8NGU1 (Q8NGU1) none fragment
887. Q8NGU3 (Q8NGU3) none Estimated / Unclassified Class A GPCRs
888. Q8NGU5 (Q8NGU5) none Estimated / Unclassified Class B GPCRs
889. Q8NGU6 (Q8NGU6) none fragment
890. Q8NGU7 (Q8NGU7) none fragment
891. Q8NGU8 (Q8NGU8) none Orphan GPCR6
892. Q8NGU9 (Q8NGU9) none Estimated / Unclassified Class A GPCRs
893. Q8NGV2 (Q8NGV2) none Estimated / Unclassified Class A GPCRs
894. Q8NGV4 (Q8NGV4) none fragment
895. Q8NGV8 (Q8NGV8) none fragment
896. Q8NGV9 (Q8NGV9) none Pheromone
897. Q8NGW0 (Q8NGW0) none Olfactory II fam 6 / MOR103-105,107-119
898. Q8NGW2 (Q8NGW2) none fragment
899. Q8NGW3 (Q8NGW3) none fragment
900. Q8NGW4 (Q8NGW4) none Estimated / Unclassified Class A GPCRs
901. Q8NGW5 (Q8NGW5) none fragment
902. Q8NGW8 (Q8NGW8) none Brain-specific angiogenesis inhibitor (BAI)
903. Q8NGW9 (Q8NGW9) none Taste receptors (T1R)
904.Q8NGX1 (Q8NGX1) none Olfactory II fam 2 / MOR256-262,270-285
905. Q8NGX4 (Q8NGX4) none fragment
906. Q8NGX7 (Q8NGX7) none fragment
907.Q8NGY0 (Q8NGY0) none Olfactory II fam 10 / MOR263-269
908. Q8NGY4 (Q8NGY4) none Olfactory II fam 6 / MOR103-105,107-119
909. Q8NGY8 (Q8NGY8) none fragment
910.Q8NGZ0 (Q8NGZ0) none Olfactory II fam 2 / MOR256-262,270-285
911. Q8NGZ7 (Q8NGZ7) none Taste receptor (T1R)
912. Q8NGZ9 (Q8NGZ9) none Olfactory II fam 2 / MOR256-262,270-285
913.Q8NH04 (Q8NH04) none Olfactory II fam 2 / MOR256-262,270-285
914. Q8NH06 (Q8NH06) none fragment
915.Q8NH07 (Q8NH07) none Olfactory II fam 11 / MOR106,121-122
916. Q8NH08 (Q8NH08) none Olfactory unclassified class II
917. Q8NH09 (Q8NH09) none Olfactory unclassified class II
918.Q8NH10 (Q8NH10) none Olfactory II fam 5 / MOR172-224,249,254
919. Q8NH11 (Q8NH11) none fragment
920. Q8NH12 (Q8NH12) none Estimated / Unclassified Class B GPCRs
921. Q8NH13 (Q8NH13) none fragment
922. Q8NH14 (Q8NH14) none Olfactory unclassified class II
923. Q8NH17 (Q8NH17) none fragment
924. Q8NH20 (Q8NH20) none fragment
925. Q8NH22 (Q8NH22) none fragment
926. Q8NH23 (Q8NH23) none fragment
927. Q8NH24 (Q8NH24) none fragment
928. Q8NH25 (Q8NH25) none fragment
929. Q8NH27 (Q8NH27) none fragment
930. Q8NH28 (Q8NH28) none fragment
931. Q8NH29 (Q8NH29) none fragment
932. Q8NH32 (Q8NH32) none fragment
933. Q8NH33 (Q8NH33) none fragment
934. Q8NH36 (Q8NH36) none Olfactory II fam 10 / MOR263-269
935. Q8NH38 (Q8NH38) none fragment
936. Q8NH39 (Q8NH39) none Purine receptor etc.
937. Q8NH44 (Q8NH44) none Olfactory Unclassified Class II
938. Q8NH45 (Q8NH45) none fragment
939. Q8NH46 (Q8NH46) none fragment
940. Q8NH47 (Q8NH47) none fragment
941. Q8NH52 (Q8NH52) none fragment
942.Q8NH58 (Q8NH58) none fragment
943. Q8NH62 (Q8NH62) none fragment
944. Q8NH65 (Q8NH65) none fragment
945. Q8NH66 (Q8NH66) none fragment
946. Q8NH68 (Q8NH68) none Olfactory I fam 51-52 / MOR1-42
947. Q8NH69 (Q8NH69) none Olfactory II fam 5 / MOR172-224,249,254
948. Q8NH71 (Q8NH71) none fragment
949. Q8NH75 (Q8NH75) none fragment
950. Q8NH77 (Q8NH77) none fragment
951.Q8NH78 (Q8NH78) none Olfactory I fam 51-52 / MOR1-42
952.Q8NH80 (Q8NH80) none Olfactory II fam 5 / MOR172-224,249,254
953.Q8NH81 (Q8NH81) none Olfactory II fam 5 / MOR172-224,249,254
954. Q8NH82 (Q8NH82) none fragment
955. Q8NH84 (Q8NH84) none Olfactory II fam 4 / MOR225-248
956. Q8NH85 (Q8NH85) none Olfactory II fam 5 / MOR172-224,249,254
957.Q8NH86 (Q8NH86) none Olfactory II fam 5 / MOR172-224,249,254
958. Q8NH88 (Q8NH88) none Olfactory II fam 5 / MOR172-224,249,254
959. Q8NH91 (Q8NH91) none Olfactory II fam 5 / MOR172-224,249,254
960. Q8NH95 (Q8NH95) none Olfactory II fam 2 / MOR256-262,270-285
961. Q8NH96 (Q8NH96) none fragment
962. Q8NH97 (Q8NH97) none fragment
963. Q8NH98 (Q8NH98) none fragment
964. Q8NH99 (Q8NH99) none Olfactory II fam 7 / MOR139-155
965. Q8NHA0 (Q8NHA0) none fragment
966. Q8NHA1 (Q8NHA1) none fragment
967. Q8NHA2 (Q8NHA2) none fragment
968. Q8NHA5 (Q8NHA5) none GABA-B Subtype 1
969. Q8NHA6 (Q8NHA6) none Olfactory II fam 2 / MOR256-262,270-285
970. Q8NHA7 (Q8NHA7) none Olfactory II fam 12 / MOR250
971. Q8NHA9 (Q8NHA9) none Metabotropic Glutamate Group III
972. Q8NHB0 (Q8NHB0) none fragment
973. Q8NHB1 (Q8NHB1) none fragment
974. Q8NHB3 (Q8NHB3) none fragment
975. Q8NHB4 (Q8NHB4) none Parathyroid hormone
976. Q8NHB5 (Q8NHB5) none Olfactory II fam 7 / MOR139-155
977. Q8NHB6 (Q8NHB6) none fragment
978. Q8NHB9 (Q8NHB9) none fragment
979. Q8NHC0 (Q8NHC0) none fragment
980. Q8NHC1 (Q8NHC1) none Olfactory II fam 7 / MOR139-155
981. Q8NHC2 (Q8NHC2) none fragment
982. Q8NHC3 (Q8NHC3) none fragment
983.Q8NHC8 (Q8NHC8) none Olfactory II fam 2 / MOR256-262,270-285
984. Q8NHD6 (Q8NHD6) none fragment
985. Q8NHZ9 (Q8NHZ9) GPRC6A Orphan GPCR6
986. Q8NI49 (Q8NI49) HRH3 Histamine Type 3
987. Q8NI50 (Q8NI50) HRH3 fragment
988. Q8TAM0 (Q8TAM0) none Purine receptor etc.
989. Q8TAN2 (Q8TAN2) none Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
990. Q8TBH9 (Q8TBH9) none Metabotropic Glutamic Acid Group II
991. Q8TBK4 (Q8TBK4) none Angiotensin type 1
992. Q8TBQ7 (Q8TBQ7) none EMR1
993. Q8TD39 (Q8TD39) none GPR37 / Endothelin B-like
994. Q8TDD6 (Q8TDD6) none Mas-related receptors (MRGs)
995. Q8TDD7 (Q8TDD7) none Mas-related receptors (MRGs)
996. Q8TDD8 (Q8TDD8) none Mas-related receptors (MRGs)
997. Q8TDD9 (Q8TDD9) none Mas-related receptors (MRGs)
998. Q8TDE0 (Q8TDE0) none Mas-related receptors (MRGs)
999. Q8TDE1 (Q8TDE1) none Mas-related receptors (MRGs)
1000. Q8TDJ8 (Q8TDJ8) none fragment
1001. Q8TDJ9 (Q8TDJ9) none Taste receptor (T1R)
1002. Q8TDP5 (Q8TDP5) CCR3 fragment
1003. Q8TDP6 (Q8TDP6) CCR3 fragment
1004. Q8TDP8 (Q8TDP8) CCR3 fragment
1005. Q8TDQ8 (Q8TDQ8) none Purine receptor etc.
1006. Q8TDS4 (Q8TDS4) GPCR Estimated / Unclassified Class A GPCRs
1007. Q8TDS5 (Q8TDS5) GPCR OR TG1019 Estimated / Unclassified Class A GPCRs
1008. Q8TDS6 (Q8TDS6) GPCR Orphan GPRC5
1009. Q8TDS7 (Q8TDS7) GPCR Mas-related receptors (MRGs)
1010. Q8TDS9 (Q8TDS9) GPCR estimation / unclassified other
1011. Q8TDT0 (Q8TDT0) GPCR estimation / unclassified, etc.
1012. Q8TDT1 (Q8TDT1) GPCR fragment
1013. Q8TDT2 (Q8TDT2) GPCR purine receptor etc.
1014. Q8TDT3 (Q8TDT3) GPCR estimation / unclassified class B GPCRs
1015. Q8TDT4 (Q8TDT4) GPCR estimation / unclassified class B GPCRs
1016. Q8TDT5 (Q8TDT5) GPCR estimation / unclassified class B GPCRs
1017. Q8TDT6 (Q8TDT6) GPCR Orphan GPCR6
1018. Q8TDT7 (Q8TDT7) GPCR estimation / unclassified, etc.
1019. Q8TDT8 (Q8TDT8) GPCR Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
1020. Q8TDT9 (Q8TDT9) GPCR Frizzled Group B (Fz 3 & 6)
1021. Q8TDU0 (Q8TDU0) GPCR estimation / unclassified, etc.
1022. Q8TDU1 (Q8TDU1) GPCR pheromone
1023. Q8TDU2 (Q8TDU2) GPCR OR V1RL4 Vomeronasal receptors V1R
1024. Q8TDU3 (Q8TDU3) GPCR vomeronasal receptors V1R
1025. Q8TDU4 (Q8TDU4) GPCR OR V1RL2 Vomeronasal receptors V1R
1026. Q8TDU5 (Q8TDU5) GPCR fragment
1027. Q8TDU6 (Q8TDU6) GPCR Estimated / Unclassified Class A GPCRs
1028. Q8TDU8 (Q8TDU8) GPCR estimation / unclassified class A GPCRs
1029. Q8TDV0 (Q8TDV0) GPCR estimation / unclassified class A GPCRs
1030. Q8TDV1 (Q8TDV1) GPCR fragment
1031. Q8TDV2 (Q8TDV2) GPCR estimation / unclassified class A GPCRs
1032. Q8TDV3 (Q8TDV3) GPCR fragment
1033. Q8TDV4 (Q8TDV4) GPCR estimation / unclassified class A GPCRs
1034. Q8TDV5 (Q8TDV5) GPCR OR GPR119 Estimated / Unclassified Class A GPCRs
1035. Q8TE23 (Q8TE23) TAS1R2 Taste receptors (T1R)
1036. Q8TEM3 (Q8TEM3) FLJ00170 fragment
1037. Q8TEV7 (Q8TEV7) MTNR1B fragment
1038. Q8WUL7 (Q8WUL7) none Melanocyte stimulating hormone
1039. Q8WUR8 (Q8WUR8) none fragment
1040. Q8WWD5 (Q8WWD5) none Prostacyclin
1041. Q8WXG9 (Q8WXG9) VLGR1 Estimated / Unclassified Class B GPCRs
1042. Q8WXR5 (Q8WXR5) CRHR1 fragment
1043. Q8WXR6 (Q8WXR6) CRHR1 fragment
1044. Q8WXR7 (Q8WXR7) CRHR1 fragment
1045. Q8WXR9 (Q8WXR9) FZD6 Frizzled group B (Fz 3 & 6)
1046. Q8WXV1 (Q8WXV1) none fragment
1047. Q8WXV2 (Q8WXV2) none fragment
1048. Q8WXZ9 (Q8WXZ9) HRH3 Histamine type 3
1049. Q8WY00 (Q8WY00) HRH3 Histamine type 3
1050. Q8WY01 (Q8WY01) HRH3 Histamine Type 3
1051. Q8WZ72 (Q8WZ72) MTNR1A fragment
1052. Q8WZ85 (Q8WZ85) PJCG2 fragment
1053. Q8WZ86 (Q8WZ86) JCG4 fragment
1054. Q8WZ87 (Q8WZ87) PJCG1 fragment
1055. Q8WZA6 (Q8WZA6) OR17-210 fragment
1056. Q93003 (Q93003) HA2AR fragment
1057.Q969F8 (Q969F8) HOT7T175 OR GPR54 GPCR 54
1058. Q96AM5 (Q96AM5) none Neuromedin U
1059. Q96B74 (Q96B74) none Frizzled Group A (Fz 1 & 2 & 4 & 5 & 7-9)
1060. Q96CD9 (Q96CD9) none fragment
1061. Q96CH1 (Q96CH1) none Estimated / Unclassified Class A GPCRs
1062. Q96DQ1 (Q96DQ1) none fragment
1063. Q96EC3 (Q96EC3) none β-adrenergic receptor type 2
1064. Q96FC5 (Q96FC5) none Rhodopsin and others
1065. Q96GE0 (P30874) none fragment
1066. Q96HB4 (Q96HB4) none Estimated / Unclassified Class B GPCRs
1067. Q96HT6 (Q96HT6) none fragment
1068. Q96IE7 (Q96IE7) none fragment
1069. Q96JW0 (Q96JW0) none Estimated / Unclassified Class B GPCRs
1070. Q96K78 (Q96K78) none Estimated / Unclassified Class B GPCRs
1071. Q96KE0 (Q96KE0) none fragment
1072. Q96KP5 (Q96KP5) CCR11 CC chemokine and others
1073. Q96LA9 (Q96LA9) MRGX4 Mas-related receptors (MRGs)
1074. Q96LB0 (Q96LB0) MRGX3 Mas-related receptors (MRGs)
1075. Q96LB1 (Q96LB1) MRGX2 OR GPCR Mas-related receptors (MRGs)
1076. Q96LB2 (Q96LB2) MRGX1 OR GPCR Mas-related receptors (MRGs)
1077. Q96LC6 (Q96LC6) CCKBR CCK Type B
1078. Q96LD9 (Q96LD9) H4 Histamine Type 4
1079. Q96MV6 (Q96MV6) none Estimated / Unclassified Class B GPCRs
1080. Q96N54 (Q96N54) none fragment
1081. Q96PE1 (Q96PE1) TEM5 OR GPR124 Estimated / Unclassified Class B GPCRs
1082.Q96QG0 (Q96QG0) AXOR12 GPCR 54
1083. Q96R43 (Q96R43) none fragment
1084. Q96R54 (Q96R54) none fragment
1085. Q96RE8 (Q96RE8) ADRA1A α-adrenergic receptor type 1
1086. Q96RG8 (Q96RG8) CHRM4 Acetylcholine Vertebrate Type 4
1087. Q96RG9 (Q96RG9) CHRM3 fragment
1088. Q96RH0 (Q96RH0) CHRM2 Acetylcholine Vertebrate Type 2
1089. Q96RH1 (Q96RH1) CHRM1 Acetylcholine Vertebrate Type 1
1090. Q96T96 (Q96T96) CCR3 fragment
1091.Q99412 (Q99412) 5-HT7 fragment
1092. Q99463 (Q99463) Y6 Neuropeptide Y type 6
1093.Q99586 (Q99586) none fragment
1094. Q99587 (Q99587) none fragment
1095. Q99642 (Q99642) none fragment
1096. Q99997 (Q99997) none fragment
1097. Q9BSP0 (Q9BSP0) none fragment
1098. Q9BX54 (Q9BX54) BAI3 fragment
1099. Q9BXA0 (Q9BXA0) none CXC chemokine type 4
1100. Q9BXA5 (Q9BXA5) GPR91 Purine receptor etc.
1101.Q9BXX6 (Q9BXX6) DRD3 fragment
1102. Q9BY15 (Q9BY15) none EMR1
1103. Q9BY61 (Q9BY61) none fragment
1104. Q9BYD7 (Q9BYD7) VTS20631 LGR-like (hormone receptors)
1105. Q9BYT4 (Q9BYT4) none fragment
1106. Q9BYX5 (Q9BYX5) CCR8 CC Chemokine Type 8
1107. Q9BYY4 (Q9NYN8) EDG1 fragment
1108. Q9BYY6 (Q9BYY6) CNR1 fragment
1109. Q9BYZ0 (Q9BYZ0) ADRB2 fragment
1110. Q9BZC5 (Q9BZC5) FKSG35 fragment
1111.Q9GZK1 (Q9GZK1) 6M1-4P * 04 OR 6M1-4P * 03 Olfactory II fam 2 / MOR256-262,270-285
1112. Q9GZK6 (Q9GZK6) 6M1-4P * 05 OR 6M1-4P * 02 Olfactory II fam 2 / MOR256-262,270-285
1113. Q9GZN0 (Q9GZN0) STRG Estimated / Unclassified Class A GPCRs
1114. Q9GZP7 (Q9GZP7) VNR19I1 OR V1RL1 OR GPCR vomeronasal receptors V1R
1115. Q9GZQ4 (Q9GZQ4) NMUR2 OR NMU2R OR TGR-1 Neuromedin U
1116. Q9H011 (Q9H011) GIR fragment
1117. Q9H1N4 (Q9H1N4) ADRA1D fragment
1118.Q9H1R0 (Q9H1R0) DJ402H5.1 fragment
1119.Q9H206 (Q9H206) none Olfactory II fam 6 / MOR103-105,107-119
1120.Q9H208 (Q9H208) none Olfactory II fam 10 / MOR263-269
1121. Q9H228 (Q9H228) GPCR lysosphingolipid & LPA (EDG)
1122. Q9H2C5 (Q9H2C5) none Olfactory I fam 51-52 / MOR1-42
1123.Q9H2C6 (Q9H2C6) none Olfactory I fam 51-52 / MOR1-42
1124. Q9H2C7 (Q9H2C7) none fragment
1125. Q9H2C8 (Q9H2C8) none Olfactory I fam 51-52 / MOR1-42
1126. Q9H2L2 (Q9H2L2) none Estimated / Unclassified Class A GPCRs
1127.Q9H339 (Q9H339) none Olfactory I fam 51-52 / MOR1-42
1128. Q9H340 (Q9H340) none Olfactory I fam 51-52 / MOR1-42
1129. Q9H342 (Q9H342) none Olfactory I fam 51-52 / MOR1-42
1130. Q9H345 (Q9H345) none Olfactory I fam 51-52 / MOR1-42
1131. Q9H573 (Q9H573) OPRM1 opioid type M
1132.Q9H7M4 (Q9H7M4) FLJ00046 fragment
1133.Q9H7Q2 (Q9H7Q2) FLJ00015 fragment
1134. Q9HAR2 (Q9HAR2) LEC3 Latrophilin Type 3
1135. Q9HAR3 (Q9HAR3) LEC2 Latrophylline type 1
1136. Q9HB44 (Q9HB44) none fragment
1137. Q9HB45 (Q9HB45) none Growth hormone-releasing hormone
1138. Q9HB89 (Q9HB89) NMUR1 Neuromedin U
1139. Q9HBV6 (Q9HBV6) HCRTR1 Orexin
1140. Q9HBW9 (Q9HBW9) ETL ETL receptors
1141. Q9HCA5 (Q9HCA5) STRL33 BONZO receptors (CXC6R)
1142. Q9NPY0 (Q9NPY0) ADRA1D fragment
1143.Q9NQ20 (Q9NQ20) GPR31 GPR
1144. Q9NQ84 (Q9NQ84) GPRC5C Orphan GPRC5
1145. Q9NQS5 (Q9NQS5) EX33 OR GPR84 OR GPCR Estimated / Unclassified Class A GPCRs
1146. Q9NRA6 (Q9GZQ4) NMU2R OR TGR-1 Neuromedin U
1147. Q9NRB8 (Q9NRB8) none Lysosphingolipid & LPA (EDG)
1148. Q9NSC9 (Q9NSC9) HPFH6OR fragment
1149. Q9NSM3 (Q9NSM3) DKFZP434B1272 fragment
1150. Q9NYK7 (Q9NYK7) none CCK Type B
1151. Q9NYN8 (Q9NYN8) CHEDG1 OR EDG1 Rhizosphingolipid & LPA (EDG)
1152. Q9NZD1 (Q9NZD1) GPRC5D Orphan GPRC5
1153. Q9NZG5 (Q9NZG5) GPRC5C Orphan GPRC5
1154. Q9NZH0 (O75205) GPRC5B Orphan GPRC5
1155. Q9NZP3 (Q9NZP3) HSA12 fragment
1156. Q9NZP4 (Q9NZP4) HSA10 fragment
1157. Q9P1R1 (Q9P1R1) none fragment
1158. Q9P1T4 (Q9P1T4) CCR5 fragment
1159. Q9P1T5 (Q9P1T5) CCR5 fragment
1160. Q9P1V4 (Q9P1V4) none fragment
1161. Q9P1Z7 (Q9P1Z7) KIAA1531 fragment
1162. Q9P2Q4 (Q9P2Q4) HTR1F fragment
1163.Q9P2Q9 (Q9P2Q9) HTR2A fragment
1164. Q9UBJ7 (Q9UBJ7) CCR5 CC Chemokine Type 5
1165. Q9UBT9 (Q9UBT9) CCR5 CC Chemokine Type 5
1166. Q9UCW0 (Q9UCW0) none fragment
1167. Q9UD23 (Q9UD23) none Endothelin
1168. Q9UD63 (Q9UD63) none α-adrenergic receptor type 1
1169. Q9UD67 (Q9UD67) none fragment
1170. Q9UDD7 (Q9UDD7) none fragment
1171. Q9UDD8 (Q9UDD8) none fragment
1172.Q9UDD9 (Q9UDD9) none Olfactory II fam 1 / MOR125-138,156
1173. Q9UDE6 (Q9UDE6) none Substance K (NK2)
1174. Q9UEB1 (Q9UEB1) none fragment
1175. Q9UHX3 (Q9UHX3) EMR2 EMR1
1176. Q9UJ42 (Q9UJ42) GPCR150 OR GPCR estimation / unclassified, etc.
1177.Q9UJ48 (Q9UJ48) LPHH1 fragment
1178.Q9UJ49 (Q9UJ49) LPHH1 fragment
1179. Q9UJ50 (Q9UJ50) LPHH1 fragment
1180. Q9UJ51 (Q9UJ51) LPHH1 fragment
1181. Q9UJ52 (Q9UJ52) LPHH1 fragment
1182. Q9UKY5 (Q9UKY5) LEC1 Latrophilin type 2
1183. Q9UKY6 (Q9UKY6) LEC1 Latrophilin Type 2
1184. Q9UL14 (Q9UL14) OR17-1 fragment
1185. Q9UL61 (Q9UL61) VLGR1 Estimated / Unclassified Class B GPCRs
1186. Q9UM77 (Q9UM77) OLFR 17-210 fragment
1187. Q9UN23 (Q9UN23) CCR5 CC Chemokine Type 5
1188. Q9UN24 (Q9UN24) CCR5 CC Chemokine Type 5
1189. Q9UN25 (Q9UN25) CCR5 CC Chemokine Type 5
1190. Q9UN26 (Q9UN26) CCR5 CC Chemokine Type 5
1191. Q9UN27 (Q9UN27) CCR5 CC Chemokine Type 5
1192. Q9UN28 (Q9UN28) CCR5 CC Chemokine Type 5
1193. Q9UPA4 (P51681) CCR5 CC Chemokine Type 5
1194. Q9UPG0 (Q9UPG0) CRAM-B CC Chemokine etc.
1195. Q9UPJ0 (Q9UPJ0) none fragment
1196. Q9UPJ1 (Q9UPJ1) none fragment
1197. Q9UQQ6 (Q9UQQ6) CCR9 CC Chemokine Type 9
1198. Q9UQS0 (Q9UQS0) none fragment
1199. Q9Y2T6 (Q9Y2T6) GPR55 Estimated / Unclassified Class A GPCRs
1200. Q9Y4B1 (Q9Y4B1) none fragment
1201. Q9Y653 (Q9Y653) GPR56 Estimated / Unclassified Class B GPCRs
1202. R3R1_HUMAN (Q9NSD7) SALPR Somatostatin- and angiogenin-like peptide
1203. R3R2_HUMAN (Q8TDU9) GPR100 Somatostatin and Angiogenin-like peptide
1204. RDC1_HUMAN (P25106) CMKOR1 OR RDC1 RDC1
1205. RGR_HUMAN (P47804) RGR Rhodopsin and others
1206. SCRC_HUMAN (P47872) SCTR Secretin
1207. SMO_HUMAN (Q99835) SMO OR SMOH Smooth
1208.SPR1_HUMAN (Q15743) GPR68 OR OGR1 GPR
1209.SRB3_HUMAN (Q9NS66) SREB3 SREB
1210. SSR1_HUMAN (P30872) SSTR1 Somatostatin Type 1
1211. SSR2_HUMAN (P30874) SSTR2 Somatostatin Type 2
1212. SSR3_HUMAN (P32745) SSTR3 Somatostatin Type 3
1213. SSR4_HUMAN (P31391) SSTR4 Somatostatin type 4
1214. SSR5_HUMAN (P35346) SSTR5 Somatostatin type 5
1215. TA2R_HUMAN (P21731) TBXA2R Thromboxane
1216. TAR1_HUMAN (Q96RJ0) TAR1 OR TA1 Trace amine
1217. TAR3_HUMAN (Q96RI9) TAR3 OR TA3 Traceamine
1218. TAR4_HUMAN (Q96RI8) TAR4 OR TA4 Traceamine
1219. TAR5_HUMAN (Q969N4) GPR102 OR TAR5 OR TA5 Traceamine
1220. TRFR_HUMAN (P34981) TRHR Thyrotropin-releasing hormone
1221. TSHR_HUMAN (P16473) TSHR Thyrotropin
1222. UR2R_HUMAN (Q9UKP6) GPR14 Urotensin II
1223. V1AR_HUMAN (P37288) AVPR1A OR AVPR1 Vasopressin
1224. V1BR_HUMAN (P47901) AVPR1B OR AVPR3 OR VPR3 Vasopressin
1225. V2R_HUMAN (P30518) AVPR2 OR ADHR OR V2R Vasopressin
1226. VIPR_HUMAN (P32241) VIPR1 Vasoactive intestinal polypeptide
1227. VIPS_HUMAN (P41587) VIPR2 OR VIP2R Vasoactive intestinal polypeptide

  The nuclear receptor is a receptor that functions as a ligand-inducible transcription control factor, transmits a ligand signal to genetic information, and controls expression of a target gene group at a transcription level. When this ligand-dependent transcription control is performed together with a basic transcription apparatus, a group of nuclear factors that interact with a receptor in a ligand binding-dependent manner has been found. These transcription coupling factors are known to form a complex without being present alone. Nuclear receptors include steroid hormone receptors and non-steroid hormone receptors. Examples of steroid hormone receptors include estradiol-17β receptor (ERα), glucocorticoid receptor (GR), and progesterone receptor. (PR), androgen receptor (AR), mineralocorticoid receptor (MR) and the like. Examples of nonsteroid hormone receptors include triiodothyronine receptor (TR), vitamin D receptor (VDR). ), Trans retinoin receptor (RAR), or PPA (peroxisome proliferator-activated) receptor.

  The cell adhesion molecule (CAM) is a protein also called an adhesion molecule, an adhesion factor, or an adhesion protein, and is a cell membrane molecule involved in adhesion between cells or between a cell and an extracellular matrix. For example, it is involved in morphogenesis, maintenance, regeneration, inflammation, tumor metastasis, etc. of multicellular tissues. Many penetrate the cell membrane and bind to the cytoskeletal system in the cell or transmit signals into the cell through the second messenger system. Examples of the cell adhesion molecule include an immunoglobulin superfamily, a cadherin superfamily, an integrin superfamily, a selectin (LECAM) family, and a MAM superfamily.

  The immunoglobulin superfamily is a molecule that contains one or more domains similar to immunoglobulins extracellularly. Examples of the immunoglobulin superfamily involved in the immune system include various immunoglobulins, major histocompatibility antigens (MHC) I and II, T cell receptors, CD2, CD3, CD4, CD8, CD28, or ICAM. Examples of the immunoglobulin superfamily involved in the nervous system include NCAM, L1 (NgCAM), NrCAM, Thy1, MAG, PO, TAG1, SC1, telecephalon, or fascyclin II and III .

  The cadherin superfamily is a molecule that was first reported as a cell membrane protein involved in cell-cell adhesion in a calcium-dependent manner, basically an extracellular domain with five intramolecular repeat structures, a transmembrane region, and A molecule comprising an intracellular domain and containing a characteristic motif of an acidic amino acid or an aromatic amino acid in the repeating structure. The cadherin superfamily includes, for example, type I (for example, N type, R type, or P type), a short type that does not include a transmembrane site and binds to the membrane via inositol, and a structure characteristic of an intracellular domain Or a proto type containing a large number of extracellular repeating structures.

  The integrin superfamily is a heterodimer of two subunits of αβ and is involved in adhesion between cells and extracellular matrix, and is closely related to cytoskeletal system in cells.

  The selectin (LECAM) family is a membrane protein having a lectin-like domain, an EGF (epidermal growth factor) -like domain, and a plurality of complement-binding-like domains extracellularly. Examples of the selectin (LECAM) family include LECAM-1 (L-selectin), LECAM-2 or LECAM-3 (P-selectin).

  The MAM superfamily is an adhesive membrane protein containing a MAM domain (named from mepurin, A5, μ), a molecule containing a characteristic amino acid sequence and forming a β sheet.

  The above-mentioned “ligand-binding protein fragment containing a ligand-binding site (ii)” that can be used as a ligand-binding protein or a derivative thereof capable of forming the complex of the present invention is a fragment of the ligand-binding protein, There is no particular limitation as long as it includes the ligand binding site of the ligand binding protein. For example, the extracellular domain in a transmembrane receptor can be mentioned.

  The “ligand-binding protein (i) or the polypeptide-containing substance (iii) containing the fragment (ii)” that can be used as a ligand-binding protein or a derivative thereof that can form the complex of the present invention is the ligand. It is not particularly limited as long as it contains a binding protein or a fragment thereof and retains the ability to bind to a ligand. For example, a fusion polypeptide containing a ligand binding protein or a fragment thereof, or a ligand binding protein or a fragment thereof And a polypeptide derivative in which a substance capable of binding to a partner substance is bound.

  In the “fusion polypeptide containing a ligand binding protein or fragment thereof”, examples of the polypeptide or peptide that can be fused with the ligand binding protein or fragment thereof include, for example, a marker sequence commonly used in gene recombination techniques and a ligand binding protein. Polypeptide (hereinafter referred to as a conjugated polypeptide) or a fragment thereof, or a polypeptide or peptide capable of binding to a partner substance.

The order in which the “ligand-binding protein or fragment thereof” and “polypeptide or peptide that can be fused to the ligand-binding protein or fragment thereof” in the fusion polypeptide are not particularly limited, but they are separated from the ligand-binding site. It is preferable to link so that “polypeptide or peptide that can be fused to a ligand-binding protein or a fragment thereof” is arranged at the position. For example, when the ligand binding protein is a transmembrane protein (for example, a transmembrane receptor or a cell adhesion molecule), the ligand binding site is usually located outside the cell, so that the polypeptide located on the cytoplasm side is used. It is preferable to link a “polypeptide or peptide that can be fused with a ligand-binding protein or a fragment thereof” to the terminal. More specifically, in the GPCR, the N-terminus is located on the outside of the cell and the C-terminus is located on the cytoplasm side. Is preferably linked to the N-terminus of the G protein or its α subunit or fragment thereof.
In these preferred embodiments, when the complex of the present invention contains a carrier and is supported on the carrier via a partner substance contained in the complex, the ligand binding site can be arranged at a position away from the carrier. The contact opportunity between the test substance and the ligand binding site can be increased.

  When preparing the polypeptide by genetic engineering, the marker sequence can be expressed by fusing with the polypeptide in-frame, if desired, to confirm the expression of the polypeptide, It is used so that confirmation or purification can be easily performed. For example, GST (glutathione-S-transferase) or a fragment thereof, FLAG epitope, histidine tag, hemagglutinin tag, myc epitope, etc. Can be mentioned.

The conjugated polypeptide can be appropriately selected depending on the ligand-binding protein used.
For example, when the ligand binding protein is a GPCR, the G protein or a subunit thereof can be used as the conjugate polypeptide. G protein includes monomeric G protein and trimeric G protein composed of α, β, and γ subunits. In the case of monomeric G protein, it is preferable to use G protein, and in the case of trimeric G protein, it is preferable to use a subunit, particularly an α subunit to which GTP and GDP bind.

  When the ligand binding protein is a GPCR and the G protein or a subunit thereof is used as the conjugate polypeptide, the combination of the GPCR and the G protein or the subunit is not particularly limited, and is a natural combination in vivo. Or a combination that can be artificially designed, but is preferably a combination in which a GPCR and a G protein or a subunit thereof can be conjugated to exert their functions, More preferably, it is a combination.

For example, when Gi is used as the G protein α subunit, GPCRs coupled to Gi include, for example, hGPCR44, catecholamine receptor α ₂ , dopamine receptor D2, muscarinic receptor m2, m4, m5, adenosine receptor A1, A3, PGE receptor EP3, leukotriene receptor BLT, Cys-LT1, sphingosine 1-phosphate receptor _{S1P 1, S1P 2, S1P 3} , or can be mentioned lysophosphatidic acid receptor LPA _1, further Among the human GPCRs described in Table 1, for example, serotonin type 1 family (numbers 1 to 5), serotonin type 2 family (numbers 6 to 8), α-adrenergic receptor type 2 family (numbers 16 to 18), CX3C chemokine Family (No. 44), cannabinoid Lee (numbers 49, 50), CXC chemokine family (numbers 52-54), CC chemokine family (numbers 60-67), histamine type 3 family (number 177), histamine type 4 family (number 178), or IL-8 Mention may be made of the receptor family (CXC1, CXC2) (numbers 180, 181). The CXC receptor family is all GPCRs coupled to Gi.

  In the present invention, it is not always necessary to use the conjugated polypeptide in a state where its function is exerted, and the entire conjugated polypeptide can be used as long as it functions as an antigen for the binding partner substance, Fragments can also be used.

  In the “polypeptide derivative in which a substance capable of binding to a partner substance is bound to a ligand-binding protein or a fragment thereof”, examples of the “substance capable of binding to a partner substance” include avidin, streptavidin, biotin, biotin / biotin Examples include acceptor complexes, lectins, sugars, antibodies or fragments thereof, antigens (for example, digoxigenin), or nucleic acids (particularly DNA). An example of each partner substance corresponding to each “substance that can bind to the partner substance” is shown in Table 2.

<Table 2>
Substances that can bind to partner substances Partner substances
Avidin or streptavidin biotin biotin, biotin / biotin acceptor complex avidin or streptavidin
Lectin sugar
Sugar lectin
Antibody or fragment thereof Antigen
Antigen antibody or fragment thereof
Nucleic acid complementary nucleic acid

  In the present invention, “ligand binding protein or derivative thereof” is preferably a fusion polypeptide of GPCR or a fragment thereof and G protein or an α subunit thereof or a fragment thereof, and a fusion of GPCR and G protein or an α subunit thereof. More preferably, a fusion polypeptide in which the C-terminal of a GPCR or a fragment thereof (particularly GPCR) and the N-terminal of a G protein or an α subunit thereof or a fragment thereof (particularly a G protein or an α subunit thereof) is linked Particularly preferred.

By expressing GPCR or a fragment thereof (particularly GPCR) in a fused form with G protein or an α subunit thereof or a fragment thereof (particularly G protein or an α subunit thereof), not only can it be used for ligand screening. It can also be used to measure the activity of a ligand or agonist / antagonist obtained by screening.
In addition, when expressed in the form of such a fusion polypeptide, GPCR or G-protein (or G protein α subunit) complex formed when GPCR is activated in cells is used. Since the fragment and the G protein or the α subunit or the fragment thereof are contained in the fusion polypeptide at a ratio of 1: 1, it is preferable in terms of maintaining the three-dimensional structure of the GPCR or the fragment.
Furthermore, in the fusion polypeptide linked at the specific end, when the complex of the present invention includes a carrier and is supported on the carrier via the partner substance contained in the complex, the ligand binding site is separated from the carrier. It can arrange | position in a distant position and can increase the contact opportunity of a to-be-tested substance and a ligand binding site.

  The second component that forms the complex of the present invention, “a partner substance that can specifically bind to a ligand-binding protein or derivative thereof at a site other than the ligand binding site” is the first component that forms the complex of the present invention. Depending on the ligand binding protein or its derivative which is a component of

For example, the ligand binding protein or derivative thereof is a polypeptide or includes at least a polypeptide portion, so any amino acid region other than the ligand binding site [eg, marker sequence, conjugated polypeptide portion, or ligand binding protein portion ( However, an antibody or an antibody fragment that can specifically bind to a ligand binding site is excluded) as the partner substance.
As the antibody, either a polyclonal antibody or a monoclonal antibody can be used. As the antibody, for example, IgG, IgA, IgE, IgM, or IgD can be used, and IgG is preferable. The antibody fragment is not particularly limited as long as it is a fragment that retains the reaction specificity of the original antibody, that is, a fragment containing an antigen recognition site. For example, Fab, Fab ′, F (ab ′) ₂ or Fv.

  In addition, when the ligand-binding protein or derivative thereof includes a “substance that can bind to a partner substance”, an appropriate partner substance should be appropriately selected according to the type of the “substance that can bind to the partner substance”. Can do. Table 2 shows examples of “substances that can bind to the partner substance” and “partner substances” corresponding thereto.

The complex of the present invention forms a complex by binding the “partner substance” part as the second component to the recognition site by the partner substance in the “ligand binding protein or derivative thereof” part as the first component. doing. The complex of the present invention is not particularly limited as long as it is a complex comprising a “ligand binding protein or derivative thereof” portion and a “partner substance” portion, but if desired, other components such as, for example, A carrier can be included.
Examples of the carrier include carriers that can be used for protein separation and purification [eg, various particles (eg, magnetic particles, metal particles, resin particles, glass particles, or ceramic particles), or chromatography carriers], or The carrier [for example, plate for ELISA, membrane, or latex particle] which can be used for immunological analysis can be mentioned.

  When the complex of the present invention includes a carrier, it is preferably supported on the carrier via a partner substance contained in the complex. For example, when the carrier is a particle, by taking such a structure, the ligand binding site of the ligand binding protein contained in the complex is arranged toward the outside of the particle. When the complex is used for screening for a substance that binds to a ligand binding protein, the chance of contact between the test substance and the ligand binding site can be increased. In addition, when the carrier is a flat plate carrier (for example, a plate or a membrane), the same effect can be obtained.

  In the complex of the present invention, the type of “ligand binding protein or derivative thereof” included as the first component is not particularly limited, but is preferably one type. When the type of the ligand binding protein or derivative thereof in the complex of the present invention is one, screening targeting only the target ligand binding protein can be performed without being affected by other ligand binding proteins. .

  In the complex of the present invention, the “ligand binding protein or derivative thereof” included as the first component is preferably an isolated ligand binding protein or derivative thereof. In the present specification, the “isolated” state means a state separated from a cell, for example, a state of a cell membrane fraction containing a ligand binding protein or a derivative thereof, or a solubilization treatment from the cell membrane fraction. Further, a purified state can be mentioned. The complex of the present invention is particularly preferably contained as a ligand-binding protein purified by solubilization treatment from a cell membrane fraction or a derivative thereof.

In a preferred embodiment of the complex of the present invention,
(1) a ligand-binding protein selected from the group consisting of a protein capable of specifically binding to a ligand, a fragment of the protein containing a ligand-binding site, and a polypeptide-containing substance containing the protein or the fragment; ,
(2) A partner substance capable of specifically binding to the ligand binding protein or derivative thereof at a site other than the ligand binding site, and substantially free of ligand binding protein or derivative thereof other than the ligand binding protein.

In another preferred embodiment of the complex of the present invention,
(1) an isolated ligand-binding protein selected from the group consisting of a protein capable of specifically binding to a ligand, a fragment of the protein containing a ligand-binding site, and a polypeptide-containing substance containing the protein or the fragment; Its derivatives and
(2) A partner substance capable of specifically binding to the ligand binding protein or derivative thereof at a site other than the ligand binding site, and substantially free of ligand binding protein or derivative thereof other than the ligand binding protein.

In a further preferred embodiment of the complex of the present invention,
(1) an isolated ligand-binding protein selected from the group consisting of a protein capable of specifically binding to a ligand, a fragment of the protein containing a ligand-binding site, and a polypeptide-containing substance containing the protein or the fragment; One of its derivatives,
(2) A partner substance capable of specifically binding to the ligand binding protein or derivative thereof at a site other than the ligand binding site, and substantially free of ligand binding protein or derivative thereof other than the ligand binding protein.

The complex of the present invention can be prepared by combining general gene recombination techniques and protein separation and purification techniques.
For example, based on a conventional method of gene recombination technology, a gene encoding a desired ligand-binding protein or derivative thereof is incorporated into an appropriate expression vector, the expression vector is transfected into an appropriate host, and the transformant is cultured. By doing so, the ligand-binding protein or a derivative thereof can be produced. Examples of the host include E. coli, yeast, mammalian cells (for example, CHO cells or COS cells), or insect cells (for example, Sf9 cells). Subsequently, the cell fraction containing the complex of the present invention is recovered by a cell fractionation method selected based on the biochemical properties of the produced ligand-binding protein or derivative thereof, and an appropriate partner substance and By contacting, the complex of the present invention can be prepared.

  For example, when the ligand binding protein is a transmembrane receptor (eg, GPCR), the receptor is produced in the host cell in a state of penetrating the cell membrane. After culturing, the cells are collected by centrifugation, the collected cells are homogenized in the presence of a protease inhibitor to disrupt the cells, and ultracentrifugated to collect the cell membrane fraction in the precipitate. As a method for disrupting cells, a known method, for example, a method of crushing cells with a homogenizer (for example, Potter-Elvehjem type), a method of disrupting with a Waring blender or polytron (manufactured by Kinematica), a method of disrupting with ultrasound, Alternatively, there may be mentioned a method of crushing cells by ejecting them from a thin nozzle while applying pressure with a French press or the like (Japanese Membrane Society, Membrane Experiment Series Volume I, Biomembrane, p. 75).

  For the cell membrane fractionation, a fractionation method using centrifugal force, for example, a fractional centrifugation method or a density gradient centrifugation method is mainly used. For example, the cell lysate is centrifuged at a low speed (500 to 3000 rpm) for a short time (usually about 1 to 10 minutes), and the supernatant is further centrifuged at a high speed (15000 to 30000 rpm) for usually 30 minutes to 2 hours. The resulting precipitate is taken as the membrane fraction. This membrane fraction is rich in the receptor protein expressed and membrane components such as cell-derived phospholipids and membrane proteins.

  The obtained cell membrane fraction is solubilized with an appropriate solubilizer (eg, digitonin, Tween 20, Lubrol PX, Triton X-100, Nonidet P-40, or CHAPS), and this is partnered with (eg, an antibody). By contacting the carrier with the carrier, a complex of the receptor and the partner substance can be obtained.

The kit of the present invention (including the screening kit and the analysis kit) comprises a “ligand binding protein is a derivative thereof” as a first component of the complex of the present invention and a “partner substance” as a second component. Each may be included as a separate substance or in the form of a complex.
Further, the kit of the present invention may be a polynucleotide containing a base sequence encoding the amino acid sequence of the ligand binding protein or derivative thereof, instead of or in addition to the “ligand binding protein or derivative thereof,” and / or A vector containing the polynucleotide can also be included.

[2] Screening method and analysis method of the present invention In the screening method of the present invention, a substance that binds to a ligand-binding protein can be screened by bringing the complex of the present invention into contact with a test substance.
The test substance that can be applied to the screening method of the present invention is not particularly limited. Or a random peptide group created by applying the phage display method or the like. In addition, culture supernatants of microorganisms, natural components or tissue extracts derived from plants, animals, or marine organisms can be used as test substances for screening. Furthermore, a compound obtained by chemically or biologically modifying a compound selected by the screening method of the present invention can be used.
By combining the screening method of the present invention and the phage display method, a peptide that binds to a target ligand-binding protein can be obtained with high probability.

The screening method of the present invention can be carried out in various embodiments depending on, for example, the state of the complex of the present invention used for screening, the type of test substance and its contact procedure, or the method used for determination. it can.
Examples of the screening method of the present invention include:
(1) a step of bringing the complex of the present invention supported on a carrier into contact with a test substance (contacting step); and (2) a step of obtaining the test substance bound to the carrier via the complex. (Acquisition process)
A screening method (hereinafter referred to as an isolated screening method), and (1) a complex of the present invention supported on a carrier (provided that the receptor is a GPCR), a test substance, In the presence of labeled GTP (contacting step), and (2) analyzing the binding of GTP to the complex (GTP analysis step)
Screening method (hereinafter referred to as GTP screening method)
Is included.

  The isolated screening method of the present invention is a method for screening a substance that binds to any ligand-binding protein. The isolated screening method of the present invention can also be used as a method for purifying a substance that binds to a ligand-binding protein.

  In the obtaining step in the isolated screening method of the present invention, the test substance bound to the carrier via the complex is separated from the unreacted test substance. For example, when using magnetic beads as the carrier for supporting the complex, magnetically, when using other beads (for example, metal beads or resin beads), by centrifugation, when using ELISA plates The separation can be performed by washing.

  When the test substance is bound to the complex supported on the carrier, the test substance can be identified and the substance that binds to the ligand binding protein can be determined. For example, when the test substance is a peptide-displayed phage, the peptide that binds to the ligand-binding protein is determined by eluting from the complex with an appropriate eluate and determining the DNA sequence encoding the displayed peptide. Can do.

  The GTP type screening method of the present invention is a method that can be applied when the receptor is a GPCR. When a ligand or an agonist binds to the GPCR, the G protein or the α subunit thereof converts from GDP to GTP. This is a method utilizing the fact that an exchange reaction occurs. According to the GTP type screening method of the present invention, not only a receptor-binding substance can be obtained, but also the action of the substance on the receptor (for example, agonistic action, antagonistic action, partial agonistic action, inverse agonistic action, or allosteric ligand) Action) can also be determined.

  In the GTP type screening method of the present invention, a complex containing a fusion polypeptide of GPCR or a fragment thereof and G protein or an α subunit or a fragment thereof is used as the complex of the present invention. In the present specification, the term “GPCR fragment” means a fragment of a GPCR and containing a ligand binding site, unless otherwise specified. The term “G protein fragment” means a G protein fragment containing a GTP binding site, unless otherwise specified, and “G protein α subunit fragment” is not particularly specified. Unless otherwise indicated, it means a fragment of the G protein α subunit and containing a GTP binding site. Hereinafter, the complex of the present invention that can be used in the GTP type screening method of the present invention is referred to as a GTP type screening complex.

In the contacting step in the GTP type screening method of the present invention, the GTP type screening complex and the test substance are contacted in the presence of labeled GTP. In addition, when screening an antagonist, it is preferable to coexist a ligand or an agonist with labeled GTP.
Examples of substances that can be used for labeling GTP include known labeling substances such as radioisotopes, luminescent substances, fluorescent substances, and enzymes.

In the GTP analysis step of the GTP type screening method of the present invention, analysis of GTP binding in the complex caused by addition of a test substance in the contact step (for example, detection of presence or absence of GTP binding or GTP binding) Measure the amount).
For example, when the test substance is an agonist, an exchange reaction from GDP to GTP occurs in the G protein or its α subunit, so that the binding amount of labeled GTP increases. Therefore, agonists can be screened using as an index the increase in the amount of labeled GTP bound when a test substance is added.

  Further, when the test substance is an antagonist, the increase in the amount of labeled GTP bound by the ligand or agonist coexisting with the labeled GTP decreases due to the action of the antagonist. Therefore, antagonists can be screened using as an index the decrease in the amount of labeled GTP bound when a test substance is added.

  When the test substance is an inverse agonist, the exchange reaction from GDP to GTP that occurs constitutively in the absence of the test substance is reduced by the action of the antagonist. Therefore, inverse agonists can be screened using the decrease in the amount of labeled GTP binding when a test substance is added as an index.

  Further, when the test substance is an allosteric ligand, it does not act antagonistically with the ligand or agonist, so that the activity depends only on the test substance concentration dependence, regardless of the ligand or agonist concentration dependence. Therefore, with the change (increase or decrease) in the amount of labeled GTP bound when the test substance is added (provided that the change does not depend on the concentration of the ligand or agonist), the allosteric ligand is Can be screened.

  In the GTP type screening method of the present invention, a mixture of a plurality of different test substances can be used as the test substance. However, in order to examine the action of each test substance, for each type of test substance, It is preferable to implement.

The analysis method of the present invention can be carried out according to the screening method of the present invention. Examples of the analysis method of the present invention include:
(1) a step of bringing the complex of the present invention supported on a carrier into contact with a test substance (contacting step); and (2) a step of obtaining the test substance bound to the carrier via the complex. (Acquisition process)
And (1) a complex of the present invention carried on a carrier (provided that the receptor is a GPCR) And a test substance in the presence of labeled GTP (contact process), and (2) a process of analyzing the binding of GTP to the complex (GTP analysis process).
A method for analyzing the binding and / or action of the test substance to the receptor (referred to as GTP type analysis method)
Is included.

  The isolated analysis method of the present invention can be carried out by the same method as the isolated screening method of the present invention. The substance obtained in the obtaining step in the isolated analysis method of the present invention is a substance that binds to the ligand-binding protein, and thus has a binding ability to the ligand-binding protein.

The GTP type analysis method of the present invention can be carried out by the same method as the GTP type screening method of the present invention.
For example, in the analysis step of the GTP type analysis method of the present invention, when an increase in the amount of labeled GTP binding is observed due to the addition of the test substance, it is determined that the test substance has an agonistic effect on the receptor. can do.
Further, when a decrease in the amount of labeled GTP binding is observed due to the addition of the test substance (for example, an increase in the amount of labeled GTP binding by a ligand or agonist coexisting with the labeled GTP is When decreased by addition), it can be determined that the test substance has an antagonistic action on the receptor.
In addition, when a decrease in the amount of labeled GTP binding is observed due to the addition of the test substance (for example, the exchange reaction from GDP to GTP that occurs constitutively in the absence of the test substance is caused by the addition of the test substance. , The test substance can be determined to have an inverse agonistic action on the receptor.
In addition, when a test substance is added, a change (increase or decrease) in the amount of labeled GTP binding is observed, and when the change does not depend on the ligand or agonist concentration, the test substance is a receptor. It can be determined to have an allosteric ligand action on

  EXAMPLES Hereinafter, the present invention will be specifically described by way of examples, but these do not limit the scope of the present invention.

Example 1: Preparation of cell membrane fraction containing human hGPCR44-Giα fusion polypeptide
In this example, a human hGPCR44-Giα fusion polypeptide in which the C-terminus of human hGPCR44 and the N-terminus of bovine Giα subunit are linked is produced in insect cell Sf9 according to the following procedure, and then the cell membrane fraction is recovered. Thus, a cell membrane fraction containing the fusion polypeptide was prepared.

  Specifically, the gene of human hGPCR44 consisting of the base sequence represented by SEQ ID NO: 1 was amplified by PCR using a primer set consisting of the base sequences represented by SEQ ID NO: 3 and SEQ ID NO: 4. The bovine Giα subunit gene consisting of the base sequence represented by SEQ ID NO: 2 was amplified by PCR using a primer set consisting of the base sequences represented by SEQ ID NO: 5 and SEQ ID NO: 6. Subsequently, by using a mixture of the amplified human hGPCR44 gene and the bovine Giα subunit gene as a template, PCR using a primer set consisting of the nucleotide sequences represented by SEQ ID NO: 3 and SEQ ID NO: 6 was carried out, whereby the human hGPCR44 gene and the bovine Giα subunit were A gene fusion gene (human hGPCR44-Giα fusion gene) was prepared. The obtained fusion gene was digested with restriction enzymes NotI and XbaI and inserted into a multicloning site (NotI and XbaI sites) of a vector pFASTBac1 (manufactured by Invitrogen) for producing recombinant baculovirus.

  Using the prepared vector, E. coli DH10Bac (manufactured by Invitrogen) retaining the baculovirus genome as Bacmid is transformed, antibiotics (kanamycin, gentamicin, and tetracycline), isopropylthiogalactoside (IPTG), and The mixture was applied to an agar medium containing Bluo-gal (Invitrogen) and cultured at 37 ° C. Among the resulting colonies, a bacmid in which homologous recombination occurred in Escherichia coli DH10Bac and the target human hGPCR44-Giα fusion gene was inserted was isolated from colonies that remained white without being blue. . In addition, each operation of the said transformation and bacmid isolation was implemented according to the attached protocol.

  An isolated recombinant bacmid (baculovirus genome) is transfected into insect cells Sf9 (Invitrogen) (moi = 1) and cultured at 28 ° C. for 96 hours, whereby a recombinant having a human hGPCR44-Giα fusion gene is obtained. Baculovirus was obtained. This recombinant virus was infected with fresh Sf9 cells and cultured at 28 ° C. for 48 hours. In addition, each operation of the said transfection and infection was implemented according to the attached protocol. During this time, the human hGPCR44-Giα fusion polypeptide is obtained in a state where the human hGPCR44-Giα fusion gene is expressed and penetrates the cell membrane. After culture, the cells were collected by centrifugation (6000 rpm, 4 ° C., 10 minutes). The collected cells were homogenized in the presence of a protease inhibitor (protease inhibitor cocktail; manufactured by Sigma), and ultracentrifugated to collect the cell membrane fraction as a precipitate. The ultracentrifugation was performed by centrifuging at a low speed (3000 rpm, 4 ° C.) for 5 minutes, and the obtained supernatant was further centrifuged at a high speed (20000 rpm, 4 ° C.) for 2.5 hours to obtain a precipitate.

<< Example 2: Screening of human hGPCR44-binding peptide from peptide-displayed phage library >>
(1) Add 50 μL of anti-G protein monoclonal antibody (Mouse Anti Gi1α Monoclonal Antibody; manufactured by SCB) (50 μg of protein amount) to an ultrafiltration tube (Centricon YM-10; manufactured by MILLIPORE), and add 100 mmol / L sodium bicarbonate buffer. 2 mL of (pH 8.0) was added and centrifuged (5000 G, 4 ° C., 60 minutes). Again, 2 mL of 100 mmol / L sodium bicarbonate buffer (pH 8.0) was added, and centrifugation (5000 G, 4 ° C., 60 minutes) was repeated twice. Finally, 100 mmol / L sodium bicarbonate buffer (pH 8.0) The volume was 500 μL.

(2) Here, 10 μL of a biotinylated reagent (EZ-Link ^™ Sulfo-NHS-LC-Bitin; manufactured by PIERCE) dissolved in 1 mL of 100 mmol / L sodium bicarbonate buffer (pH 8.0) was added and lightly added. After stirring, the mixture was incubated at 4 ° C. for 8 hours. After 20 mL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl) was passed through a gel filtration column (Hi Trap Desulting 5 × 5; manufactured by Amersham), the entire amount of the incubated mixture was allowed to flow. Immediately, 1 mL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl) was allowed to flow, followed by 2 mL. The biotinylated antibody was obtained by recovering the entire amount of liquid discharged from the column when flowing 2 mL.

(3) Put 100 μL of magnetic beads (Dynabeads M-280 streptavidin; manufactured by Dynal) with streptavidin immobilized on the surface in a 1.5 mL tube, and add 1 mL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl). Then, the washing operation of separating by magnetism and discarding the buffer was performed 6 times. The biotinylated antibody 1mL collect | recovered by said (2) was added here, and it stirred at room temperature for 1 hour with Aicle (Aikuru RBM-100; Asahi Techno Glass). Then, the solution is separated by magnetism and the solution is discarded. After washing 5 times with 1 mL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl), 1 mL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl) is added. In addition, the obtained antibody-immobilized magnetic bead suspension was stored at 4 ° C.

(4) The human hGPCR44-Giα fusion polypeptide expression membrane (10 mg / mL) prepared in Example 1 stored at −20 ° C. was gently thawed on ice, and 500 μL thereof was put into a 1.5 mL tube. The mixture was centrifuged at 12000 rpm for 1 minute in a tabletop centrifuge (Diskboy FB-4000; manufactured by KURABO). Discard the supernatant and add 1 mL of 0.4% digitonin buffer [0.1 mol / L HEPES, 10 mmol / L MgCl ₂ , 160 mmol / L NaCl, 1 mmol / L EDTA, 0.4% digitonin (SERVA), pH 8.0] and pipette. And suspended in ice for 2 hours. Centrifugation was performed at 12000 rpm for 1 minute in a tabletop centrifuge.

(5) Take 250 μL of the antibody-immobilized magnetic bead suspension prepared in (3) above in a 1.5 mL tube, and add 0.1% in TBS-T buffer [TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl). Washed 6 times with the addition of% Tween20. To this, the whole amount of the supernatant (solubilized human hGPCR44-Giα fusion polypeptide) centrifuged in (4) above was added and incubated in ice for 2 hours.

(6) Separately from (5), 250 μL of the antibody-immobilized magnetic bead suspension prepared in (3) above was taken in a 1.5 mL tube and washed 6 times with TBS-T buffer. A random peptide-displayed phage library (0.01 g of skim milk added to 1 mL of TBS-T buffer and 20 μL of library) was added thereto, and the mixture was stirred at room temperature for 1 hour or more with an eyeicle. As the library, a library prepared based on the description of Smith, GP, Science, 288, 1315-1317 (1985) and JK Scott and GP Smith, Science, 249, 386-390 (1990) is used. did.

(7) During the incubation in (6) above, the magnetic beads were magnetically separated from the magnetic bead suspension incubated in (5) above, and then the contents were discarded and washed 3 times with TBS-T buffer. . To this, 1 mL of blocking buffer (2% skim milk / TBS-T buffer) was added and incubated for 1 hour in ice.

(8) The magnetic beads (solubilized human hGPCR44-Giα fusion polypeptide is immobilized on the magnetic beads from the magnetic bead suspension incubated in (7) above via the antibody immobilized on the magnetic beads. ) Is separated magnetically from the magnetic bead suspension of (6) above, and the supernatant (containing a random peptide-displayed phage library from which substances adsorbed to the antibody-immobilized magnetic beads are removed) is contained here. added. Incubate for 1 hour in ice. After washing 10 times with TBS-T, 100 μL of elution buffer [0.2 mol / L glycine-HCl, 0.1% bovine serum albumin (BSA) pH 2.2] was added and incubated at room temperature for 7 minutes. After magnetic beads are separated by magnetism, the supernatant is added to a 0.5 mL tube to which 15 μL of neutralization buffer (1 mol / L Tris-HCl pH 0.1) has been added, and stirred gently to elute phage solution. Got.

(9) Into a 300 mL Erlenmeyer flask was placed 20 mL of LB medium [10 g of bactotryptone, 5 g of yeast extract, and 5 g of NaCl dissolved in 1 L of pure water (Milli-Q water)], and then overnight in LB medium. An appropriate amount of E. coli (ER2738; manufactured by BioLabs), which had been cultured at 37 ° C. with shaking, was added and cultured at 37 ° C. with shaking. When the OD ₆₀₀ reached about 0.6, the entire amount of the eluted phage solution prepared in the above (8) was added and cultured with shaking at 37 ° C. for 4.5 hours. After completion of the culture, the whole amount was transferred to a 50 mL centrifuge tube, and centrifuged for 10 minutes at 4 ° C. and 10,000 rpm with a centrifuge (himac CR22F; manufactured by Hitachi, Ltd.). The supernatant was transferred to a 50 mL centrifuge tube containing 4 mL of PEG solution (20% PEG # 6000, 2.5 mol / L NaCl) and incubated at 4 ° C. for an hour. Centrifugation was performed at 10,000 rpm and 4 ° C. for 15 minutes, the supernatant was discarded, and the precipitate was suspended in 1 mL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl). This was transferred to a 1.5 mL tube, centrifuged at 10000 rpm, 4 ° C. for 5 minutes, and the supernatant was 1.5 mL in which 200 μL of PEG solution (20% PEG # 6000, 2.5 mol / L NaCl) was placed. Transfer to a tube and incubate in ice for 1 hour. Centrifugation was performed at 10,000 rpm and 4 ° C. for 10 minutes, the supernatant was discarded, and the precipitate was suspended in 200 μL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl) and transferred to a 0.5 mL tube.

(10) The steps (4) to (9) were repeated three times. However, the third time was up to (8). During this repetition, the phage used in (9) was used as the phage used in (6). As the amount used, the second time was 200 μL of the total amount obtained in the first time, the third time and the fourth time were 100 μL, which was half of the amount obtained in the second time and the third time, respectively. In addition, after transferring the supernatant in the third and fourth (6) steps, phages were eluted from the beads by the same method as in (8). This is a peptide-displayed phage bound to an antibody.

(11) The fourth target binding phage solution and the antibody binding phage solution were panned into single clones with a titer according to the following procedure. The phage solution was diluted to 10 2 to 10 4 with TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl). Distribute 200 μL of E. coli that has been cultured overnight in LB medium at 37 ° C. into a 1.5 mL tube, dispense 10 μL of the diluted phage solution, incubate for 2.5 minutes, and dissolve in an autoclave. is allowed agarose top had been [Bacto tryptone 5g, yeast extract 2.5g, NaCl2.5g, agarose (1600) 3.5 g, and pure water _{MgCl 2 · 6H 2 O 0.5g (} Milli-Q water) After being dissolved in 500 mL, autoclaved], autoclaved by adding 15 g of agar to 1 L of LB medium, added with IPTG 0.05 g, X-gal 0.04 g, and 1 mL of dimethylformamide, and placed in a sterile petri dish ). After solidification, it was sealed with parafilm and incubated at 37 ° C. for about 8 hours. A plate with around 100 plaques was selected, E. coli cultured at an OD ₆₀₀ of about 0.6 was dispensed into a test tube at a rate of 5 mL, the plaque on the plate was struck with a toothpick, and the toothpick was placed in a test tube. This operation was performed on 12 plaques on each plate. After shaking culture at 37 ° C. for 4.5 hours, the whole amount was transferred to a 50 mL centrifuge tube and centrifuged at 10,000 rpm and 4 ° C. for 10 minutes. The supernatant was transferred to a 50 mL centrifuge tube containing 1 mL of PEG solution (20% PEG # 6000, 2.5 mol / L NaCl) and incubated at 4 ° C. for 8 hours. Centrifugation was performed at 10,000 rpm and 4 ° C. for 15 minutes, the supernatant was discarded, and the precipitate was suspended in 1 mL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl). This was transferred to a 1.5 mL tube, centrifuged at 10000 rpm, 4 ° C. for 5 minutes, and the supernatant was 1.5 mL in which 200 μL of PEG solution (20% PEG # 6000, 2.5 mol / L NaCl) was placed. Transfer to a tube and incubate in ice for 1 hour. Centrifugation was performed at 10,000 rpm and 4 ° C. for 10 minutes, the supernatant was discarded, and the precipitate was suspended in 200 μL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl) and transferred to a 0.5 mL tube.

(12) The same anti-G protein monoclonal used in (1) on a 96-well plate [8 columns (A column to H column) × 12 rows (1 row to 12 rows), flat bottom high binding type; manufactured by Corning) A solution in which 50 μg of antibody was dissolved in 1.2 mL of 100 mmol / L sodium bicarbonate buffer (pH 8.0) was dispensed. An appropriate amount of 100 mmol / L sodium bicarbonate buffer (pH 8.0) was added so that the total amount per hole was 100 μL. In addition, an antibody-containing 100 mmol / L sodium bicarbonate buffer (pH 8.0) was added to the B row and the D row, and as an antibody-immobilized well, a 100 mmol / L sodium bicarbonate buffer ( Only pH 8.0) was added to serve as control wells. This was incubated at 4 ° C. for 8 hours. The inner solution was discarded, and the blocking buffer (NaCl 8 g, KCl 0.2 g, Na ₂ HPO ₄ 1.44 g, and KH ₂ PO ₄ 0.24 g was dissolved in 1 L of water and adjusted to pH 7.4 with skim milk. Was added to a concentration of 2%), and 300 μL per well was added, and the mixture was stirred with a plate stirrer at room temperature for 3 hours. The contents were discarded and washed 5 times with 200 μL per well of a washing buffer (0.5 mL of Tween 20 added to 1 L of PBS buffer).

  A solution obtained by adding 20 μL of phage eluate bound to the antibody to 80 μL of blocking buffer was added to each clone in a well. In addition, using the combination of row A (antibody-immobilized well) and row B (control well) or the combination of row C (antibody-immobilized well) and row D (control well), add the same clone, A total of 20 clones were added. The plate was stirred for 1 hour at room temperature. The contents were discarded and washed 5 times with 200 μL per well of washing buffer. HRP (horseradish peroxidase) labeled antibody (HRP / Anti M13 Monoclonal conjugate; manufactured by amersham pharmacia biotech) 20 μL against the phage used was added to 10 mL of blocking buffer, and 100 μL per well was added. The plate was stirred for 1 hour at room temperature. The contents were discarded and washed 6 times with 200 μL per well of washing buffer. A substrate (ABTS2 ammonium salt; manufactured by Wako Pure Chemical Industries, Ltd.) (1.7 mg) was dissolved in 0.05 mol / L citric acid solution (7.5 mL), and 30% hydrogen peroxide solution (13 μL) was added thereto. After 30 minutes, the absorbance was measured with a plate reader (ARVO SX; manufactured by Wallac). The absorbance of each clone (No. 1 to No. 20) is shown in Table 3, and the ratio of the absorbance in the antibody-immobilized well to the absorbance in the control well (“absorbance in the antibody-immobilized well” / “absorbance in the control well”) Table 4 shows.

<< Table 3 >>
No. 1 2 3 4 5 6 7 8 9 10
Row A 0.117 0.086 0.093 0.064 0.092 0.061 0.078 0.071 0.069 0.077
Row B 0.607 0.466 0.591 0.584 0.590 0.432 0.714 0.670 0.681 0.805

No. 11 12 13 14 15 16 17 18 19 20
Column C 0.094 0.075 0.092 0.081 0.073 0.068 0.073 0.082 0.078 0.048
Row D 0.673 0.497 0.774 0.624 0.526 0.540 0.656 0.658 0.650 0.047

<< Table 4 >>
No. 1 2 3 4 5 6 7 8 9 10
5.196 5.386 6.372 9.184 6.406 7.106 9.173 9.403 9.879 10.446

No. 11 12 13 14 15 16 17 18 19 20
7.155 6.617 8.413 7.677 7.238 7.938 8.965 8.027 8.321 0.972

(13) When the ratio in Table 4 is 1 or more, it means that the peptide-displayed phage is bound to the antibody, so the DNA of the clone was sequenced according to the following procedure. 30 μL of the phage solution (11) was placed in a 0.5 mL tube, 6 μL of a PEG solution (20% PEG # 6000, 2.5 mol / L NaCl) was added, and the mixture was incubated on ice for 1 hour. Centrifuge at 10000 rpm, 4 ° C. for 10 minutes, discard the supernatant and discard the precipitate (Iodide) buffer (10 mmol / L Tris-HCl, 1 mmol / L EDTA ₂ Na, 4 mol / L NaI, pH 8.0) It melt | dissolved in 100 microliters, 250 microliters of 100% ethanol was added there, and it incubated for 10 minutes. Centrifugation was performed at 11000 rpm and 4 ° C. for 10 minutes, the supernatant was discarded, and 300 μL of 70% ethanol was added to gently wash the inside of the tube. After drying with a vacuum dryer (CENTRIFUGAL VAPORIZER CVE-100; manufactured by EEYLA), 30 μL of sterilized pure water (Milli-Q water) was added and dissolved well. A 0.5 mL tube was newly prepared, and sterilized pure water (Milli-Q water) 12 μL, phage DNA solution 5 μL, and primer (SEQ ID NO: 6) solution (5 pmol / L) 1 μL were added and stirred. 4 μL each was dispensed into 0.2 mL color tubes (green, blue, yellow, and red). A fluorescently labeled primer kit (Thermo Sequenase fluorescent labeled primer cycle sequencing kid; manufactured by amersham pharmacia biotech) was added according to the attached protocol, and 95 ° C. (30 seconds) with a thermal cycler (TaKaRa PCR Thermal Cycler; manufactured by TaKaRa BIOMEDICALS). A PCR reaction comprising 30 cycles of 55 ° C. (20 seconds) and 70 ° C. (60 seconds) was performed. 5 μL of electrophoresis dye (included in Thermo Sequenase fluorescent labeled primer cycle sequencing kid; manufactured by amersham pharmacia biotech) was added, and phage DNA was sequenced using a DNA sequencer (manufactured by Amersham). As the amino acid sequence of the peptide encoded from the obtained base sequence, clone no. From 1, 5, 6, 8, 13, 14, and 16, the same amino acid sequence consisting of 7 amino acid residues was obtained. In addition, clone no. From 12 and 15, the same amino acid sequence consisting of 7 amino acid residues different from the amino acid sequence was obtained. Furthermore, clone no. From 2, 7, 9, 10 and 11, different amino acid sequences each consisting of 7 amino acid residues were obtained. In addition, clone No. 3 and 4 cannot determine the nucleotide sequence, and clone no. 17-20 did not perform base sequence determination.

(14) The target-binding phage was also subjected to DNA sequencing by the same method as (13). However, since no ELISA was performed, all clones were subjected to DNA sequencing. As the amino acid sequence of the peptide encoded from the obtained base sequence, clone no. From 2 and 10, the same amino acid sequence consisting of 7 amino acid residues (molecular weight calculated from the amino acid sequence = 969 Da) was obtained. From 1, 3 to 9, 11, and 12, the same amino acid sequence consisting of 12 amino acid residues (molecular weight calculated from the amino acid sequence = 1702 Da) was obtained. Since the peptide sequence bound to the target human hGPCR44 is different from any of the antibody-bound peptide sequences, it was found that a peptide sequence with specificity was obtained. Hereinafter, clone no. 2 and 10 are referred to as clone A and clone no. 1, 3-9, 11, and 12 are referred to as clone B.

Example 3: Measurement of activity of peptide-displayed phage that binds to human hGPCR44
(1) For clones A and B obtained in Example 2, phages were amplified by the following procedure. 20 mL of LB medium was placed in a 300 mL Erlenmeyer flask, and an appropriate amount of E. coli (ER2738) that had been cultured with shaking in LB medium at 37 ° C. overnight was added thereto, followed by incubation with shaking at 37 ° C. When the OD ₆₀₀ reached about 0.6, the toothpick was immersed in the phage solution cloned in (11) of Example 2, and the toothpick was put into an Erlenmeyer flask and cultured with shaking at 37 ° C. for 4.5 hours. When the cultivation was completed, the whole amount was transferred to a 50 mL centrifuge tube, and centrifuged for 10 minutes at 10,000 rpm and 4 ° C. in a centrifuge. The supernatant was transferred to a 50 mL centrifuge tube containing 4 mL of PEG solution (20% PEG # 6000, 2.5 mol / L NaCl) and incubated at 4 ° C. for 8 hours. Centrifugation was performed at 10,000 rpm and 4 ° C. for 15 minutes, the supernatant was discarded, and the precipitate was suspended in 1 mL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl). This was transferred to a 1.5 mL tube, centrifuged at 10000 rpm, 4 ° C. for 5 minutes, and the supernatant was a 1.5 mL tube containing 200 μL of PEG solution (20% PEG # 6000, 2.5 mol / L NaCl). And incubated in ice for 1 hour. Centrifugation was performed at 10,000 rpm and 4 ° C. for 10 minutes, the supernatant was discarded, and the precipitate was suspended in 20 μL of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl) and transferred to a 0.5 mL tube.

(2) Reaction solution [final concentration: 20 mmol / L HEPES buffer (pH 7.0), 1 mmol / L EDTA, 160 mmol / L NaCl, 1 mmol / L dithiothreitol (DTT), 10 mmol / L MgCl ₂ , 1 μmol / L GDP, 0.2 mg / mL A cell membrane containing human hGPCR44-Giα fusion polypeptide, and 100 pmol / L [ ³⁵ S] GTPγS] in 92 μL, clone A or B (10 ¹⁴ pfu / 4 mL of 20 mL / L peptide and 4 μL of a known agonist (peptide consisting of the amino acid sequence represented by SEQ ID NO: 8, 10 nmol / L or 50 nmol / L) was added, and the total amount was 100 μL at 30 ° C. for 30 minutes. Reacted.
In addition, as a control, phage and a known agonist were not added (that is, reaction solution only), and TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl) and only a known agonist were added (that is, phage was not added, which was known) Reaction with addition of agonist only) was performed. Further, as a comparative example, a phage (clone No. 10) displaying a peptide that binds to an antibody was reacted with a known agonist.

Thereafter, a filter (GF / B filter; manufactured by Perkin Elmer) was set in a harvester and filtered to collect [ ³⁵ S] GTPγS bound to the receptor. The filter is thoroughly washed with 30 mmol / L phosphate buffer, dried at 60 ° C. for 15 minutes, then placed in a vial, scintillation cocktail is added, and [ ³⁵ S] GTPγS remaining on the filter is removed with a liquid scintillation counter. It was measured.

  The results are shown in FIG. In FIG. 1, lanes 1 to 4 are when the known agonist is 10 nmol / L, lanes 5 to 8 are when the known agonist is 50 nmol / L, and lane 9 is when no phage and no known agonist are added. Is the result of Lanes 1 and 5 are the results of clone A in the presence of a known agonist, lanes 2 and 6 are results of clone B in the presence of a known agonist, and lanes 3 and 7 are clones in the presence of a known agonist. No. The results are 10 and lanes 4 and 8 are the results of addition of TBS buffer (50 mmol / L Tris-HCl, 150 mmol / L NaCl) and a known agonist.

  As is clear from FIG. 1, the count amount (lane 9) observed as a background even in the absence of the known agonist increased in a dose-dependent manner by adding the known agonist (lanes 4 and 8). The increase in the count by this known agonist did not change significantly even when phage N0.10 displaying a peptide that binds to the antibody was added (lanes 3 and 7), whereas it bound to human hGPCR44. When clone A or B, which is a peptide-displaying phage, was added, a significant decrease was observed (lanes 1, 2, 5, and 6). These results indicate that phage clones A and B are antagonists, respectively, and it was found that the screening system of the present invention is useful for screening substances that bind to the receptor.

(3) In the above (2), the action of human hGPCR44-binding peptides was evaluated in the state of peptide-presenting phage. In order to further confirm the action of these peptides, a peptide presented by phage clone B was synthesized, A similar experiment was performed using a synthetic peptide (12mer). However, the concentration of the known agonist (peptide consisting of the amino acid sequence represented by SEQ ID NO: 8) was 10 nmol / L, and the amount of synthetic peptide added was changed. The results are shown in FIG.

In FIG. 2, the arrow indicates the concentration of a known agonist (10 ⁻⁸ mol / L). In the results shown in FIG. 2, the count when no known agonist was added was 6220.6 cpm, and the count when only the known agonist was added (ie, no synthetic peptide was added) was 8439.3 cpm (FIG. 2). Not shown).
As shown in FIG. 2, it was found that when the synthetic peptide was added stepwise from 10 ⁻⁹ mol / L, the activity of a known agonist was decreased in a concentration-dependent manner, and the function as an antagonist was confirmed. Indicated.

  The complex of the present invention can be applied to use for screening a substance that binds to a ligand-binding protein.

It is a graph which shows the result of the activity measurement of the peptide display phage which couple | bonds with human hGPCR44. It is a graph which shows the result of activity measurement of a synthetic peptide.

  The numerical heading <223> in the sequence listing below describes the “Artificial Sequence”. Each base sequence represented by the sequence of SEQ ID NO: 3 to 7 is an artificially synthesized primer sequence, and the base sequence represented by the sequence of SEQ ID NO: 8 is a known agonist of hGPCR44.

Claims (15)

(1) selected from the group consisting of (i) a protein capable of specifically binding to a ligand, (ii) a fragment of the protein containing a ligand binding site, and (iii) a polypeptide-containing substance containing the protein or the fragment A ligand binding protein or derivative thereof;
(2) A complex comprising a partner substance capable of specifically binding to the ligand-binding protein or a derivative thereof at a site other than the ligand-binding site.   The complex according to claim 1, wherein the ligand-binding protein or a derivative thereof is a G protein-coupled receptor, a fragment of the receptor containing a ligand binding site, or a polypeptide-containing substance containing the receptor or the fragment. body.   The complex according to claim 1, wherein the ligand-binding protein or derivative thereof is hGPCR44, a fragment of the receptor containing a ligand-binding site, or a polypeptide-containing substance containing the receptor or the fragment.   The complex according to claim 1, wherein the ligand-binding protein or a derivative thereof is a fusion polypeptide of a G protein-coupled receptor or a fragment thereof and a G protein or an α subunit or a fragment thereof.   The complex according to claim 1, wherein the ligand-binding protein or derivative thereof is a fusion polypeptide of hGPCR44 or a fragment thereof and G protein or an α subunit or a fragment thereof.   The complex according to any one of claims 1 to 5, wherein the partner substance is an antibody.   The complex according to any one of claims 1 to 5, wherein the partner substance is an IgG antibody.   A screening method for a substance that binds to a ligand-binding protein, comprising contacting the complex according to any one of claims 1 to 7 with a test substance.   The method according to claim 8, wherein the ligand-binding protein or derivative thereof in the complex is a fusion polypeptide of a G protein-coupled receptor or a fragment thereof and a G protein or an α subunit thereof or a fragment thereof. (1) (1a) a ligand-binding protein selected from the group consisting of a protein capable of specifically binding to a ligand, a fragment of the protein containing a ligand-binding site, and a polypeptide-containing substance containing the protein or the fragment, or a derivative thereof (1b) a polynucleotide comprising a base sequence encoding the amino acid sequence of the ligand-binding protein or derivative thereof, or (1c) a vector comprising the polynucleotide;
(2) A kit for screening a substance that binds to the ligand-binding protein, comprising a partner substance that can specifically bind to the ligand-binding protein or a derivative thereof at a site other than the ligand-binding site.   The kit according to claim 10, wherein the ligand-binding protein or a derivative thereof is a fusion polypeptide of a G protein-coupled receptor or a fragment thereof and a G protein or an α subunit thereof or a fragment thereof.   A method for analyzing the binding and / or action of the test substance to the ligand-binding protein, wherein the test substance is brought into contact with the complex according to claim 1.   The method according to claim 12, wherein the ligand-binding protein or derivative thereof in the complex is a fusion polypeptide of a G protein-coupled receptor or a fragment thereof and a G protein or an α subunit thereof or a fragment thereof. (1) (1a) a ligand-binding protein selected from the group consisting of a protein capable of specifically binding to a ligand, a fragment of the protein containing a ligand-binding site, and a polypeptide-containing substance containing the protein or the fragment, or a derivative thereof (1b) a polynucleotide comprising a base sequence encoding the amino acid sequence of the ligand-binding protein or derivative thereof, or (1c) a vector comprising the polynucleotide;
(2) a binding and / or action of a test substance to the ligand binding protein, comprising a partner substance capable of specifically binding to the ligand binding protein or a derivative thereof at a site other than the ligand binding site Analysis kit.   The kit according to claim 14, wherein the ligand-binding protein or a derivative thereof is a fusion polypeptide of a G protein-coupled receptor or a fragment thereof and a G protein or an α subunit thereof or a fragment thereof.

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