[go: up one dir, main page]

WO1993003137A1 - Recepteur de substance p humaine - Google Patents

Recepteur de substance p humaine Download PDF

Info

Publication number
WO1993003137A1
WO1993003137A1 PCT/US1992/006532 US9206532W WO9303137A1 WO 1993003137 A1 WO1993003137 A1 WO 1993003137A1 US 9206532 W US9206532 W US 9206532W WO 9303137 A1 WO9303137 A1 WO 9303137A1
Authority
WO
WIPO (PCT)
Prior art keywords
val
ser
leu
tyr
ala
Prior art date
Application number
PCT/US1992/006532
Other languages
English (en)
Inventor
James E. Krause
Original Assignee
Washington University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Washington University filed Critical Washington University
Publication of WO1993003137A1 publication Critical patent/WO1993003137A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70571—Receptors; Cell surface antigens; Cell surface determinants for neuromediators, e.g. serotonin receptor, dopamine receptor

Definitions

  • This invention relates to the human substance P receptor. More particularly, the invention concerns the molecular cloning and functional expression of the human substance P receptor and a novel stable cell line that expresses large numbers of the recombinant substance P receptor.
  • Substance P is a peptide neurotransmitter and neuromodulator originally detected in 1931 based on its smooth muscle contractile activity (1) .
  • SP was isolated based on its sialagogic activity, and its primary structure was established as Arg-Pro-Lys-Pro- Glu-Gln-Phe-Phe-Gly-Leu-Met-NH 2 (2) [SEQ ID NO:l].
  • SP has since been shown to participate in the regulation of diverse biological activities (3,4,5), and it is an excitatory agent released from central, peripheral and gastrointestinal neurons.
  • SP regulates certain endocrine and exocrine gland secretions, it aids in the regulation of blood pressure by acting at both central and peripheral sites, and it has been suggested to be involved in the regulation of some immunological disorders and certain inflammatory states.
  • the biological actions of SP are mediated largely via a receptor that interacts specifically with the conserved tachykinin carboxyl terminal domain.
  • the specific amino terminal sequences of the mammalian tachykinin peptides dictate receptor affinity and selectivity.
  • Ligand interaction with the SPR activates guanyl nucleotide binding protein dependent second messenger systems that mediate the specific biological response.
  • Yokota et al. (6) and Hershey and Krau ⁇ e (7) molecularly characterized and functionally expressed the rat SP receptor (SPR) , and established it to be a member of the G-protein coupled receptor superfamily.
  • a cDNA encoding the novel human substance P receptor has been isolated and characterized, and nucleotide sequence analysis has been used to deduce the primary structure of the receptor protein.
  • the human substance P receptor consists of 407 amino acid residues and is a member of the G-protein coupled receptor superfamily. Comparison of the novel human and the prior art rat substance P receptor amino acid sequences demonstrated that they have a 94.5% identity that is largely evident in transmembrane domains, and in intracellular domains.
  • the novel human substance P receptor was transiently expressed from plasmid pl ⁇ hSPR in a COS-7 cell line and showed a Kd for Tyr ⁇ -substance P binding of 0.24 nM.
  • a clonal cell line stably expressing the novel human substance P receptor from plasmid p- ⁇ hSPR was created in a CHO cell background, said cell line being designated herein as CHO-pM ⁇ -hSPR #10. This cell line expresses 500,000 substance P receptors per cell with an affinity of 0.29 nM.
  • a culture of this cell line is on deposit under the Budapest Treaty with the American Type Culture Collection, Rockville, MD, under accession number ATCC CRL 10824.
  • CHO cells Choinese hamster ovary
  • COS-7 cells monkey kidney, SV40 transformed
  • murine cells HeLa cells
  • canine cells and the like can similarly be used as host cells for expressiqn of the substance P receptor.
  • the pattern of ligand displacement by naturally occurring tachykinin peptide was substance P»neurokinin A>neurokinin B.
  • Ligand stimulation of transfected cells results in a rapid and transient inositol 1,4,5- triphosphate response.
  • RNA blot hybridization and solution hybridization demonstrated that the naturally expressed human substance P receptor mRNA was about 4.5 Kb in size, and was expressed in IM-9 lymphoblast and U373-MG astrocytoma cells, as well as in spinal cord and lung but not in liver.
  • Cell lines containing the human substance P receptor cDNA are useful for examining cellular mechanisms regulating human substance P receptor mRNA expression and for screening for antagonists of human substance P such as may be useful for central, peripheral and gastrointestinal system disorders, inflammation and immune disorders. Since tissues in the human body that express the substance P receptor express only about 5-10,000 receptors per cell, the substantially and significantly higher expression system of about 500,000 receptors per cell in accordance with the present invention permits rapid and faster screening of candidate compounds acting at the substance P receptor.
  • the human substance P receptor also is useful as a diagnostic approach for identifying abberrant receptor sequences in human disease states.
  • FIG. 1 shows the nucleotide sequence and deduced amino acid sequence [SEQ ID NO:3] of the human substance P receptor. Nucleotide numbering shown on the right side starts with +1 beginning with A of the initiator methionine codon. .Amino acid sequence is numbered below the displayed sequence. The putative ⁇ -helical transmembrane domains labeled MI-MVII are underlined.
  • FIG. 2 shows the expression of the human substance P receptor in COS-7 cells in two bar graph panels.
  • A Comparison of 125 l-Tyr "1 -SP binding to nontransfected cells and to cells transfected with a plasmid encoding either the human SPR, the human SPR in the antisense orientation or the rat SPR.
  • B Competition of 125 I-Tyr "t -SP binding by naturally occurring and synthetic tachykinin peptides. Transfection conditions and ligand binding were performed with 0.1 nM 125 I-Tyr "1 -SP as described in Methods hereinbelow. Each datum represents the X ⁇ SEM of four duplicate determinations performed with different preparations of transfected cells.
  • FIG. 3 is a graphical representation which shows the displacement of 125 I-Tyr "1 -SP binding to transfected C0S-7 cells by the naturally occurring tachykinins: substance P, neurokinin A and neurokinin B. Transfection conditions and ligand binding were performed with 0.1 nM 15 I-Tyr "1 -SP as described in Methods hereinbelow. Each datum point represents the X ⁇ of four determination's performed in duplicate. The SEM was less than 5% for all data presented.
  • FIG. 4 is a graphical representation which shows the saturation analysis of 125 I-Tyr "1 -SP binding to transfected COS-7 cells. Cells were transfected and ligand binding was performed as described in Methods hereinbelow.
  • the data shown are from four determinations performed in duplicate on separate transfected cell preparations.
  • the variation in ligand concentration for all data points was less than 3% of the mean concentration shown.
  • the calculated Kd and ⁇ mx values were 0.24 ⁇ and 151,000 ⁇ per cell, respectively.
  • FIG. 5 is a graphical representation which shows a human substance P stimulated inositol trisphosphate response as a function of time after stimulation of transfected COS-7 cells.
  • Cells were transfected, harvested and stimulated with 1 ⁇ M human substance P, and inositol trisphosphate levels were determined as described in Methods hereinbelow.
  • the data shown represents the results from a single transfection and stimulation test. Similar results were obtained in a repeat of the test.
  • FIG. 6 shows the analysis of human substance P receptor mRNA expression patterns by RNA blot and solution hybridization methods.
  • the upper left shows the RNA blot results
  • the upper right shows the solution hybridization-nuclease protection results
  • the lower portion illustrates the probes used.
  • RNA blots 2 ⁇ g poly(A) * RNA was denatured, electrophoresed on 1% gels and transferred to nitrocellulose as described in
  • RNA blot were 0.24 to 9.5 Kb RNA ladder (BRL, Gaithersburg, MD) , and standards for the nuclease protection gel were radiolabeled Mspl-digested pBR322.
  • FIG. 7 shows a comparison of the amino acid sequences of human substance P receptor [SEQ ID NO:3] and rat substance P receptor [SEQ ID NO:7]. Identical residues between the two sequences are indicated by the vertical line. Putative membrane spanning domains MI- MVII are overlined. The closed triangles indicate consensus N-linked glycosylation sites, the filled circles indicate potential intracellular serine and threonine phosphorylation sites, and the arrow depicts a potential palmitoylation site.
  • nucleotide bases are designated as adenine (A) ; thymine (T) ; guanine (G) ; and cytosine (C) .
  • Corresponding nucleotides are, for example, deoxyadenosine-5 « -triphosphate (dATP) .
  • Amino acids are shown either by conventional three or one letter abbreviations as follows:
  • IM-9 immunoblast cells were obtained from Drs. Norman Boyd and Susan Leeman, University of Massachusetts Medical Center, and U373 MG astrocyte cells were obtained from the ATCC (ATCC HTB 17) .
  • Tyr "1 - Substance P (Tyr-Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly- Leu-Met-NH 2 ) [SEQ ID NO:4] was synthesized by the Washington University Protein Chemistry Facility and was purified to homogeneity by HPLC using the general procedures previously described (11) . Radioiodination of peptide was performed using the conventional chloramine T oxidative iodination procedure and HPLC purification of the monoiodo form of Tyr "1 -SP.
  • RNA isolation, cDNA and ⁇ enomic cloning PCR methods and nucleotide sequence analysis. The methods for RNA isolation, (poly(A) * RNA selection and cDNA synthesis are conventional and have been described previously (7, 8, 9). PCR was performed using a Perkin Elmer thermal cycler as previously described (7) with IM-9 cDNA as target. Initially a cDNA was generated by PCR using oligonucleotide primers corresponding to G- protein coupled receptor membrane spanning domains II and VII..
  • a 671 bp cDNA was isolated, subcloned into BLUESCRIPT (pBS) and sequenced it contained an open reading frame with 90.5% identity to the corresponding rat substance P receptor cDNA (6,7) and gene (9) sequence.
  • This plasmid was termed pBS-hSPRII-VII and the inserted cDNA corresponded to nucleotides 237-908 of that shown in Figure 1.
  • the 5' and 3' extents of the hSPR cDNA coding region as well as nontranslated sequences were determined by isolation and characterization of human SPR genomic exons 1 and 5, respectively, using the pBS-hSPR II-VII cDNA insert and rat genomic exons (9) as probes, and is shown in Figure 1.
  • the predicted coding region of the human SPR was generated by PCR with IM-9 cDNA by using oligonucleotides corresponding to the coding region 5' (5'CCACCATGGATAACGTCCTCCCGGTG 3 « ) [SEQ ID NO:5] and 3' (antisense, 5•CTAGGAGAGCACATTGGAGGAGAA3') [SEQ ID NO:6] ends as primers.
  • the cDNA generated was isolated by agarose gel electrophoresis and was blunt-end ligated into Smal-digested pBS. Electroporation of bacterial cells with the ligated DNA yielded multiple isolates that were further analyzed by restriction mapping and by nucleotide sequence analysis.
  • cDNA (corresponding to bases -5 to +1227 of that shown in Fig. 1) was isolated after restriction with Hindlll and BamHI (present in the pBS polylinker) , and was made blunt- ended with Klenow fragment.
  • the pM 2 was also blunt- ended with Klenow after BamHI digestion.
  • the cDNA was ligated to pM 2 , and was used to transform E. coli XL-1 Blue cells by electroporation. Colonies containing inserts were identified and the orientation of inserts was determined by restriction analysis. Two plasmids, called pM 2 -hSPR and pM 2 -hSPR antisense, were identified. Sequence analysis was performed as described previously (7, 8, 9).
  • RNA blot and solution hybridizations were performed by conventional procedures as described previously (8, 9, 16, 17).
  • a random-primer labeled cDNA was prepared with Klenow fragment of DNA polymerase I for the pBS-hSPRII-VII cDNA insert, and an antisense RNA was prepared by transcription using T 7 RNA polymerase and EcoRl linearized pBS-hSPRII-VII.
  • RNA gels (1.0%) were blotted onto Nytran membranes, and the protected RNA species from solution hybridization tests were electrophoresed on 6% polyacrylamide gels containing 7M urea. Autoradiography was performed at -70 * with an intensifying screen.
  • a human SPR cDNA fragment corresponding to nucleotides +237 to +908 in Figure 1 was generated by PCR from cDNA prepared from IM-9 lymphoblast cell RNA using conventional procedures previously described (7) .
  • the 5' end of the coding region was determined by isolation and sequence analysis of the human SPR gene exon 1
  • the 3• end of the cDNA was determined by isolation and sequence analysis of the human SPR gene exon 5 as described in Methods hereinbefore.
  • These sequences provided the 5' and 3* translated sequences of the human SPR, and a PCR using IM-9 cell cDNA was used to generate a full coding region containing cDNA.
  • This cDNA was subcloned into the pM 2 expression vector in which the cDNA is under the control of the Harvey murine sarcoma virus LTR (10) and was used for functional expression.
  • COS-7 cells were transfected with pM 2 -hSPR, pM 2 hSPR antisense and pM ⁇ SPR, three plasmids that contain the human SPR cDNA, the human SPR cDNA inserted in the antisense orientation and the rat SPR cDNA (7) , and 48 to 72 hours later the cells were examined for binding of 125 I-Tyr *1 -SP u ing a rapid filtration assay.
  • Figure 2A shows these results in which the human and rat SPR construct transfected cells bind 15,000 to 25,000 cpm ligand that is displaced by 1 ⁇ M SP. Nontransfected cells or cells transfected with pM ⁇ SPR antisense showed no specific binding.
  • FIG. 2B shows that at 10 nM SP or physalaemin, specific 125 I- Tyr- *1 -SP binding was reduced by 85 to 95%, whereas 10- fold higher concentrations of tachykinins potent at NK- 2 and NK-3 receptors, including neurokinin A, neurokinin B, neuropeptide ⁇ , neuropeptide K, eledoisin and senktide, were much less potent in displacing radiolabelled ligand binding.
  • substance P free acid was much less potent in this regard, thereby demonstrating the importance of the substance P carboxyamide moiety in ligand binding.
  • Additional tests were performed with SP, NKA and NKB at various doses to determine the IC 50 values for displacing 125 I-Tyr "1 -SP binding, and these data are shown in Figure 3.
  • SP was the most potent displacer of ligand binding compared to NKA and NKB, with IC 50 values of 0.72 ⁇ 0.9 nM, 0.63 ⁇ 0.06 ⁇ M, and 1.12 ⁇ 0.21 ⁇ M, respectively, each with Hill coefficients of 0.94 - 0.96.
  • this cloned human cDNA encodes a sequence in transfected cells that upon ligand binding analysis has the characteristics of a SPR or NK-1 type tachykinin binding site.
  • Saturation analysis of 125 l-Tyr *1 -SP binding was performed to determine the affinity and relative number of binding sites expressed by pH ⁇ hSPR transfection of COS-7 cells.
  • Figure 4 shows these results; S ⁇ atchard analysis of this data by the ligand program (12) provides a 1 site fit with a k d value of 0.24 ⁇ 0.01 nM, with an average of 151,000 ⁇ 8,000 sites expressed per cell.
  • Transiently transfected COS-7 cells were stimulated with 1 ⁇ M SP to determine cellular inositol- 1,4,5 trisphosphate responses. Two tests were performed in which time points after stimulation of 5, 10, 15, 20, 30, 60 and 120 seconds were analyzed. A transient response of 2.5 to 3-fold above resting levels was observed (Figure 5) at 10 to 15 seconds after stimulation with a return to basal level by 20 to 30 seconds.
  • RNA isolated from cell lines or tissues Some patterns of human SPR RNA expression were also examined using RNA isolated from cell lines or tissues, and these data are shown in Figure 6.
  • RNA isolated from cell lines or tissues By northern blot analysis, a single hybridizing species of approximately 4.5 kb was identified in poly(A) * RNA isolated from IM-9 cells but not in liver. Similar hybridizing species were observed with both an RNA coding region probe and a DNA probe corresponding to exon I of the SPR gene.
  • the coding region antisense RNA probe was also used in a more sensitive solution hybridization-nuclease protection test in which the probe corresponds to a 712 base sequence which when annealed with hSPR mRNA, will protect a species of 671 bases.
  • Figure 6 shows that this probe will protect a species of 671 bases in IM-9 and U373 cell RNA preparation and also in spinal cord and lung RNA preparations, that correspond to SPR mRNA; these are not observed in HepG2 cell or liver RNA preparations.
  • IM-9 cell RNA preparations two additional protected species of approximately 150 bases and 350 bases were noted. These species have not been completely characterized, but they may correspond to exon 1 protected species and exons 1 plus 2 protected species which would be present in partially spliced RNAs. These have been observed in rat tissues (9) and appear to correspond to slowly spliced nuclear SPR RNA precursors.
  • Figure 7 shows a comparison of primary structures of human and rat SPR protein, as deduced from cDNA cloning and sequence analysis. Twenty-two of the 407 residues are different between the sequences; these differences are distributed throughout and are generally conservative. Both sequences encode receptors with 7 putative ⁇ -helical transmembrane domains based on hydrophobicity plotting and by comparisons to other G- protein coupled receptors.
  • the human SPR has 2 N-linked glycosylation sites in the amino terminal domain, and a potential pal itoylation site (cys-323) 15 residues carboxyl terminal to the MVII transmembrane domain.
  • the carboxyl terminal tail region is separated by an acidic region about half way into the sequence, which separates the two Ser/Thr rich regions.
  • MOLECULE TYPE protein
  • MOLECULE TYPE protein

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Cell Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Neurology (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

L'invention concerne l'isolement, la caractérisation et le codage par ADNc du récepteur de substance P humaine, la structure primaire du récepteur protéique, ainsi qu'une lignée céllulaire CHO transformée à l'aide d'un vecteur d'expression d'ADN contenant un ADNc codant le récepteur de substance P humaine.
PCT/US1992/006532 1991-08-07 1992-08-05 Recepteur de substance p humaine WO1993003137A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US74120091A 1991-08-07 1991-08-07
US741,200 1991-08-07

Publications (1)

Publication Number Publication Date
WO1993003137A1 true WO1993003137A1 (fr) 1993-02-18

Family

ID=24979773

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1992/006532 WO1993003137A1 (fr) 1991-08-07 1992-08-05 Recepteur de substance p humaine

Country Status (2)

Country Link
AU (1) AU2444692A (fr)
WO (1) WO1993003137A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0830600A4 (fr) * 1995-06-07 2000-04-05 Praecis Pharm Inc Dosage biologique fonctionnel pour les agonistes et antagonistes des recepteurs couples de proteine g
US6114139A (en) * 1994-08-11 2000-09-05 Takeda Chemical Industries, Ltd. G-protein coupled receptor protein and a DNA encoding the receptor
EP1100962A1 (fr) * 1998-07-25 2001-05-23 AstraZeneca AB Polymorphismes genetiques du gene du recepteur neurokininique 1 de l'homme et leurs utilisations pour le diagnostic et le traitement d'affections
GB2370274A (en) * 2000-12-19 2002-06-26 Warner Lambert Co Modified Tachykinin receptors
WO2002013799A3 (fr) * 2000-08-18 2003-03-13 Univ Mcgill Oligonucleotides et autres modulateurs de la voie du recepteur nk-1 et utilisations therapeutiques de ces derniers
WO2005090401A1 (fr) * 2004-03-20 2005-09-29 Astrazeneca Ab Molecules

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BIOCHEMISTRY, Volume 30, No. 44, issued 1991, N.P. GERARD et al., "Human Substance P Receptor(NK-1): Organization of the Gene, Chromosome Localization, and Functional Expression of cDNA Clones", pages 10640-10646. *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6114139A (en) * 1994-08-11 2000-09-05 Takeda Chemical Industries, Ltd. G-protein coupled receptor protein and a DNA encoding the receptor
EP0830600A4 (fr) * 1995-06-07 2000-04-05 Praecis Pharm Inc Dosage biologique fonctionnel pour les agonistes et antagonistes des recepteurs couples de proteine g
EP1100962A1 (fr) * 1998-07-25 2001-05-23 AstraZeneca AB Polymorphismes genetiques du gene du recepteur neurokininique 1 de l'homme et leurs utilisations pour le diagnostic et le traitement d'affections
WO2002013799A3 (fr) * 2000-08-18 2003-03-13 Univ Mcgill Oligonucleotides et autres modulateurs de la voie du recepteur nk-1 et utilisations therapeutiques de ces derniers
GB2370274A (en) * 2000-12-19 2002-06-26 Warner Lambert Co Modified Tachykinin receptors
WO2005090401A1 (fr) * 2004-03-20 2005-09-29 Astrazeneca Ab Molecules

Also Published As

Publication number Publication date
AU2444692A (en) 1993-03-02

Similar Documents

Publication Publication Date Title
Takeda et al. Molecular cloning, structural characterization and functional expression of the human substance P receptor
Lachowicz et al. Molecular cloning of a novel G protein‐coupled receptor related to the opiate receptor family
Marchese et al. Cloning of human genes encoding novel G protein-coupled receptors
Wank et al. Brain and gastrointestinal cholecystokinin receptor family: structure and functional expression.
EP0746332B2 (fr) Adn codant un recepteur de polypeptide pancreatique (y4) et utilisation de celui-ci
US5889151A (en) Purified human alpha interferon receptor
EP1015478A1 (fr) Criblage de bibliotheques servant de strategie pour cloner des medicaments agissant sur les recepteurs couples a la proteine g
FR2642075A1 (fr) Polypeptides ayant une activite de recepteur (beta)-adrenergique chez l'homme, impliques dans la reponse lipolytique, acides nucleiques codant pour ces polypeptides et utilisation de ces polypeptides pour le criblage de substance active sur ces polypeptides
Chan et al. P-glycoprotein genes in the winter flounder, Pleuronectes americanus: isolation of two types of genomic clones carrying 3′ terminal exons
WO1997041263A9 (fr) Detection du recepteur de la leptine dans des organes reproducteurs et procedes de regulation de la biologie reproductrice
CA2222939A1 (fr) Peptides et composes se fixant sur le recepteur il-1
US20040152875A1 (en) Novel purinergic receptor
EP0656007A1 (fr) Genes de recepteurs opioides
WO2001098330A2 (fr) Lignee cellulaire recombinee exprimant gpcrx11 en tant que recepteur fonctionnel valide par l'angiopeptine et utile dans le criblage d'agonistes et antagonistes
WO1993003137A1 (fr) Recepteur de substance p humaine
US6447996B1 (en) Galanin receptors, nucleic acids, transformed cells and uses thereof
EP0804452A1 (fr) Recepteur c140 recombinant et ses agonistes et antagonistes
EP0906424A2 (fr) Recepteurs mixtes mammaliens de lymphocytes et de chimoquine
WO2002059349A2 (fr) Recepteurs d'odorants et leur utilisation
US20020086359A1 (en) Novel guanosine triphosphate (GTP) binding protein-coupled receptor proteins
US6416972B1 (en) DNA encoding a prostaglandin F2α receptor, a host cell transformed therewith and an expression product thereof
WO1992010565A1 (fr) Recepteur d'hormone de liberation de thyrotropine pituitaire
US20030092035A1 (en) Pain signaling molecules
JPH10507904A (ja) ヒトのカッパオピオイド受容体、核酸と使用
CA2139431A1 (fr) Polypeptides ayant une activite de recepteur serotoninergique (5ht6), acides nucleiques codant pour ces polypeptides et utilisations

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL SE

NENP Non-entry into the national phase

Ref country code: CA

122 Ep: pct application non-entry in european phase