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WO1994028019A1 - Capteur de calcium chez l'homme - Google Patents

Capteur de calcium chez l'homme Download PDF

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Publication number
WO1994028019A1
WO1994028019A1 PCT/SE1994/000483 SE9400483W WO9428019A1 WO 1994028019 A1 WO1994028019 A1 WO 1994028019A1 SE 9400483 W SE9400483 W SE 9400483W WO 9428019 A1 WO9428019 A1 WO 9428019A1
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WO
WIPO (PCT)
Prior art keywords
calcium
leu
aaa
gac
protein
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PCT/SE1994/000483
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English (en)
Inventor
Göran ÅKERSTRÖM
Lars Klareskog
Claes Juhlin
Lars Rask
Original Assignee
Aakerstroem Goeran
Lars Klareskog
Claes Juhlin
Lars Rask
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 Aakerstroem Goeran, Lars Klareskog, Claes Juhlin, Lars Rask filed Critical Aakerstroem Goeran
Priority to EP94917235A priority Critical patent/EP0700401A1/fr
Priority to AU69021/94A priority patent/AU685127B2/en
Priority to JP6525857A priority patent/JPH08510135A/ja
Publication of WO1994028019A1 publication Critical patent/WO1994028019A1/fr
Priority to US08/476,515 priority patent/US6239270B1/en
Priority to US08/652,877 priority patent/US6187548B1/en

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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/02—Nutrients, e.g. vitamins, minerals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/12—Drugs for disorders of the metabolism for electrolyte homeostasis
    • A61P3/14—Drugs for disorders of the metabolism for electrolyte homeostasis for calcium homeostasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to a cDNA clone encoding a human calcium sensor protein of parathyroid, placental, and kidney tubule cells.
  • WO 88/03271 there is described monoclonal antiparathyroid antibodies identifying a parathyroid cell membrane-bound calcium receptor or sensor, crucially involved in calcium regulation of the parathyroid hormone (PTH) release (1,2).
  • the receptor function is essential for maintenance of normal plasma calcium concentrations, and reduced receptor expression within proliferating parathyroid cells of patients with hyperparathyroidism (HPT) results in calcium insensitivity of the PTH secretion and variably severe hypercalcemia (3-6).
  • Reactivity with the antiparathyroid antibodies was also demonstrated for proximal kidney tubule cells and cytotrophoblast cells of the human placenta, and the cytotrophoblasts were demonstrated to exhibit an almost parathyroid-identical regulation of cytoplasmic calcium [Ca 2+ i] (7,8).
  • the antibody-reactive structure was found to exert calcium sensing function also in the cytotrophoblasts, and as these cells constitute part of the syncytium, the calcium sensor was suggested to be actively involved in the calcium homeostasis of the fetus (7,8). It was proposed that the antibody-reactive structure of the proximal kidney tubule cells exerts a similar calcium sensing function, and that the calcium sensor, thus, plays a more universal role in calcium regulation via different organ systems (1,7,9,10).
  • an object of the present invention was to provide sufficient structural data of the calcium sensor/receptor to enable complete characterization thereof.
  • the present invention provides probes for identifying other novel calcium sensor protein.
  • Another object was to use said structural data to design novel treatment methods as well as compounds and compositions for treating calcium related disorders.
  • cells expressing the calcium sensor protein or comprising the cDNA encoding the calcium sensor protein of the present invention may be utilized in an assay to identify molecules which block or enhance the activity of the calcium sensor protein. These molecules will be useful in the treatment of mammalian pathological conditions associated with perturbations in the levels of PTH, vitamin, D3 production, estrogen, osteoclast activity or osteoblast activity (therefore, bone resorption and/or formation), calcium secretion and calcium ion homeostasis.
  • the present invention describes the isolation and partial characterization of a cDNA clone encoding the calcium sensor/-receptor in human placenta and Northern blots verifying the presence of the corresponding mRNA within the parathyroid and kidney.
  • Close sequence similarity between the calcium sensor and a previously described rat Heymann nephritis antigen (11) suggests that the common calcium sensor of the placenta, the parathyroid and kidney tubule is related to this antigen, or represents its human homologue, and that it belongs to a family of large glycoproteins with receptor function and calcium binding ability.
  • Fig 1 Isolation by HPLC of peptides obtained after digestion of the calcium-sensor protein with Lys-C endoprotease (solid line). Dashed line represents the chromatography of an identical reaction where the calcium-sensor was omitted. The flow rate was kept at 100 ⁇ l/min. Two peptide fractions which gave easily interpretable sequences are denoted by arrows.
  • Fig 3 Partial nucleotide sequence (SEQ ID No. 3) and deduced amino acid sequence (SEQ ID No. 4) of the cDNA clone, pCAS-2, encoding part of the calcium-sensor protein. Portions of the deduced amino acid sequence identical to the peptides 292 and 293 are underlined.
  • Fig 4. Alignment of the amino acid sequence of the calcium-sensor protein (SEQ ID No. 4) to corresponding portions of the Heymann antigen (HEYMANN, SEQ ID No. 5), low density lipoprotein receptor (LDL-RC, SEQ ID No. 6), and LDL related receptor protein (LDL-RRP, SEQ ID No. 7). Stars denote residues identical between the calcium sensor protein and any of the other sequences. X denotes a position in the Heymann antigen sequence where identity has not been published.
  • polypeptide means a linear array of amino acids connected one to the other by peptide bonds between the ⁇ -amino and carboxy groups of adjacent amino acids.
  • substantially purified is used herein to mean “substantially homogeneous”, which is defined as a material which is substantially free of compounds normally associated with it in its natural state (e.g., other proteins or peptides, carbohydrates, lipids). "Substantially purified” is not meant to exclude artificial or synthetic mixtures with other compounds. The term is also not meant to exclude the presence of impurities which do not interfere with biological activity, and which may be present, for example, due to incomplete purification or compounding with a pharmaceutically acceptable preparation.
  • biologically active polypeptide means the naturally occurring polypeptide per se as well as biologically active analogues thereof, including synthetically produced polypeptides and analogues thereof, as well as natural and pharmaceutically acceptable salts and pharmaceutically acceptable derivatives thereof.
  • biologically active polypeptide also encompasses biologically active fragments thereof, as well as “biologically active sequence analogues” thereof. Different forms of the peptide may exist. These variations may be characterized by difference in the nucleotide sequence of the structural gene coding for proteins of identical biological function.
  • biologically active sequence analogue includes non-naturally occurring analogues having single or multiple amino acid substitutions, deletions, additions, or replacements. All such allelic variations, modifications, and analogues resulting in derivatives which retain one or more of the native biologically active properties are included within the scope of this invention.
  • nucleotides are indicated by their bases using the following standard one-letter abbreviations:
  • amino acid residues are indicated using the following standard one-letter abbreviations:
  • amino acid as used herein is meant to denote the above recited natural amino acids and functional equivalents thereof.
  • This invention provides an isolated nucleic acid molecule encoding the calcium sensor protein and having a coding sequence comprising the sequence shown in Fig. 3 (SEQ ID No. 3).
  • this invention provides a vector comprising an isolated nucleic acid molecule encoding the calcium sensor protein.
  • the invention provides a method of preparing calcium sensor protein which comprises inserting a nuleic acid encoding calcium sensor in a suitable vector, inserting the resuting vector in a suitable host cell, recovering the calcium sensor protein produced by the resulting cell, and puryfying the calcium sensor protein so recovered.
  • This method for prepararing calcium sensor protein uses recombinant DNA technology methods which are well known in the art.
  • the present invention also provides antisense nucleic acids which can be used to down regulate or block the expression of the calcium sensor protein either in vitro, ex vivo or in vivo.
  • the down regulation of gene expression can be made at both translational or transcriptional levels.
  • Antisense nucleic acids of the invention are more preferentially RNA fragments capable of specifically hybridizing with all or part of the sequence SEQ ID No. 3 or the corresponding messenger RNA.
  • These antisense can be synthetic oligonucleotides prepared based on the sequence SEQ ID No. 3, optionally modified to improve their stability of selectivity, as disclosed for instance in EP 92574. They can also be DNA sequences whose expression in the cell produces RNA complementary to all or part of the calcium sensor protein mRNA.
  • These antisenses can be prepared expression of all or part of the sequence SEQ ID No. 3 in the opposite orientation (EP 140 308).
  • Tissue specimens Samples of human parathyroid glands were obtained at surgery of patients with primary HPT. Other human tissue specimens (kidney, epididymis, liver, pancreas, adrenal gland, small gut, spleen, lung and striated muscle) were sampled from organs removed at surgery. Human placental tissue was collected in conjunction with uncomplicated pregnancies at full term. All specimens were immediately quick-frozen in isopentane and stored at -70°C.
  • 500 kDa calcium sensor protein was isolated and purified, from altogether 25 human placentas, by immunosorbent and ion exchange chromatographies, following a previously described protocol (7).
  • the procedure utilizes two different monoclonal antiparathyroid antibodies (1,7), E11 and G11, known to bind different epitopes of the calcium sensing protein; Ell has displayed no functional effect, while G11 efficiently blocks calcium regulation in both parathyroid and placental cells (1,7).
  • the calcium sensor protein preparation was subjected to gel chromatography on a Zorbax GF25 gel column (9.2 ⁇ 250 mm), prior to enzymatic digestion.
  • the biologically active calcium sensor protein of the present invention has been isolated as described. It can also be preparared by chemical synthesis of in a recombinant DNA biosystem. Biologically active fragments of the calcium sensor protein can also be prepared using synthetic or recombinant technologies which are known in the art.
  • Oligonucleotide synthesis Oligonucleotides were synthesized using an ABI 381 oligonucleotide synthesizer (Applied Biosystems). The following oligonucleotide mixture was utilized as a probe for screening of the placental cDNA library:
  • the first nine nucleotides contain an EcoR I and a BamH I site, respectively, and the remaining nucleotides correspond to amino acid residues 1 to 6 of peptide 293 and to residues 9 to 14 of peptide 292.
  • a placental ⁇ gt 11 cDNA library (Clontech, Ca., USA) was plated out to a density of approximately 2 ⁇ 10 5 plaques within a 20x25 cm agar plate.
  • PCR reaction Part of the ⁇ gt 11 cDNA clone CAS-1 was amplified by PCR using two degenerated probes corresponding to portions of peptides 292 and 293. The following conditions were used: 170 ng template DNA, 1 pmol of each oligonucleotide mixture as primers, dNTP 3mM, Taq-polymerase 0.75 u. The reaction was carried out in 20 ⁇ l of 10mM Tris-HCl, pH 8.0, 1.5 mM MgCl 2 , 50mM KCl in a Perkin-Elmer 9600 PCR-machine (Perkin-Elmer, Norwalk, USA).
  • a placental ⁇ ZAP-II cDNA library was screened with the PCR-fragment from the cDNA clone CAS-1 labeled by random priming as the probe. The screening was carried out as above. 2 ⁇ 10 6 plaques distributed on ten 20 ⁇ 25 cm agar plates were screened.
  • Nucleotide sequence determination The insert of the phage clone CAS-2 was released from the phage vector in the Bluescript+ vector using a helper phage (Stratagene, La Jolla, Ca.). Nucleotide sequence reactions were carried out according to the cycle sequencing procedure, utilizing a kit from Applied Biosystems. Sequences were analyzed in an ABI 373 A DNA sequenator using the Data Collection Program VIII soft-ware (Applied Biosystems).
  • For Northern blot analysis approximately 10 ⁇ g of total RNA was electrophoresed in a 1.5%/37% agarose/formaldehyde gel, blotted onto nylon membranes (Qiabrane, Diagen GmbH, D ⁇ sseldorf, Germany) and probed with the 2.3 kb clone (see results) labeled by the random priming method.
  • Hybridizations were performed at 42°C for 18-24 h in 50% formamide, 4 ⁇ saline sodium citrate (SSC; 300 mM NaCl, 30 mM Na-citrate, pH 7.0), 2 ⁇ Denhart's solution, 10% dextran sulfate (Kabi-Pharmacia, Uppsala, Sweden) and 100 ⁇ g/ml salmon sperm DNA. Filters were washed at a final stringency of 1 ⁇ SSC/0.1% SDS for 30 min at 42°C, and exposed within a phosphorimager as above.
  • the calcium sensor protein was purified from placental tissue by means of lectin chromatography, immunosorbent chromatography utilizing the immobilized monoclonal anti-parathyroid antibodies, and finally ion exchange chromatography (1,7).
  • the same antibodies were used in a sandwich ELISA to monitor the purification (7).
  • the whole final preparation consisting of 200 ⁇ g of the 500 kDa protein chain (7), was made 6 M with regard to guanidine-HCl and applied to a gel chromatography column, equilibrated with 2 M guanidine-HCl, 0.1 M Tris-Cl, pH 8.5. The column was eluted with the same buffer.
  • An oligonucleotide mixture (48 bp) was constructed to encode amino acid residues 2 to 17 of the sequenced peptide 292.
  • five inosine bases were inserted at degenerated positions where no guidance could be obtained from the codon usage in humans. At nine positions, where two bases were possible, one of the bases was suggested with a likelihood exceeding 70% from codon usage, and was therefore used in the oligonucleotide mixture.
  • the mixed oligonucleotide was radioactively labelled and used as a probe to screen a human placental ⁇ gt 11 cDNA library. Approximately 2 ⁇ 10 6 plaques were screened and a single positive clone, CAS-1, was found. The insert of this clone was estimated to 2.3 kb, by restriction mapping. To obtain a recognizable sequence of the clone in a rapid way, an attempt was made to PCR amplify part of the sequence using degenerated oliogonucleotides corresponding to part of peptides 292 and 293 as primers. A distinct DNA fragment of approximately 430 bp was obtained assuming that the peptide 292 is located carboxy-terminal to peptide 293.
  • the fragment was partially sequenced using the oligonucleotide mixture corresponding to peptide 293 as the primer. In one reading frame from the obtained sequence, the sequence VGRHI could be deduced, in excellent agreement with the carboxyterminal 5 amino residues of peptide 293.
  • a human placental ⁇ ZAP-II cDNA library reported to contain clones with large inserts was screened with the PCR-fragment as the probe. From 2 ⁇ 10 6 plaques a single clone, CAS-2, was found. The insert of this clone, estimated to 2.8 kb, was released in the Bluescript + vector, using a helper phage.
  • the 500 kDa placental calcium sensor belongs to the LDL-receptor superfamily.
  • Fig 4 shows an alignment of the available placental 500 kDa protein sequence to the sequence of the Heymann antigen (SEQ ID No. 5) as well as to two other members of the same protein superfamily, the LDL-receptor (SEQ ID No. 6) and the LDL-receptor-related protein (identical to the ⁇ 2 -macroglobulin receptor, 11,15,16, SEQ ID No. 7).
  • the antiparathyroid antibodies (E11 and G11) were found to stain not only parathyroid, placental and proximal kidney tubule cells but also epididymal cells, as previously demonstrated for antibodies reactive with the Heymann antigen (17-20).
  • RNA total RNA (approximately 10 ⁇ g/lane) from human kidney, placenta and parathyroid glands with the identified 2.8 kb clone as the probe, revealed one major hybridizing RNA species of approximately 15 000 bases in all these tissues (Fig 5). Human liver, pancreas, adrenal gland, and small gut (Fig 5) as well as spleen, lung and striated muscle (not shown) lacked hybridizing species.
  • parathyroid As key regulator of the calcium homeostasis has been related to its extraordinarily capacity to sense and respond to variation in the extracellular Ca 2+ ion concentration.
  • Essential for recognition of changes in external calcium is a cation receptor or sensor of the parathyroid cell membrane, the presence of which was implicated by a series of in vitro studies on parathyroid cell regulation (9,10,21-24).
  • the concept of a cell membrane receptor was further, substantiated when monoclonal antiparathyroid antibodies were found to recognize and interfere with the calcium sensing of parathyroid cells (1-6).
  • cytotrophoblast cells of the human placenta selected by their reactivity with the antiparathyroid antibodies, displayed parathyroid-like sensing of changes in external calcium, a function which also could be blocked by one of the anti-parathyroid antibodies (7,8).
  • the calcium sensor of the placenta was subsequently isolated by immunosorbent and ion exchange chromatographies and shown to consist of a large glycoprotein of approximately 500 kDa molecular size (7). It was also demonstrated by immunoprecipitation that a protein of the same size reacted with the antiparathyroid antibodies within the parathyroid and kidney tubule cells (to be published, (25).
  • the parathyroid calcium sensor or receptor is known to have features in common with most other classical receptors for cellular activation, although it exhibits the unusual ability to bind and be activated by divalent cations. Cation binding triggers biphasic rise in [Ca 2+ i] and concomittant activation of phospholipase C, possibly via a coupled G-protein, with a resulting accumulation of inositol phosphates (2,5,9,10).
  • the LDL- receptor the LDL-receptor-related protein and the Heymann antigen
  • the Ca 2+ binding is necessary for the interaction of the LDL-receptor with apo-B (27).
  • the LDL-receptor related protein ( ⁇ 2 -macroglobulin receptor) is also known to bind Ca 2+ , which induces conformational changes, and Ca 2* is necessary for binding of activated ⁇ 2 -macroglobulin to the receptor (16).
  • the rat Heymann antigen was shown by a blotting technique to interact with Ca 2+ (28).
  • the Ca 2+ binding motifs of the calcium sensor protein yet remain to be identified.
  • the sensor protein (as well as the Heymann antigen) contains EGF-like modules, like other members of the LDL-receptor superfamily (11,16,27), which may represent putative Ca 2+ binding sites.
  • EGF-like modules like other members of the LDL-receptor superfamily (11,16,27), which may represent putative Ca 2+ binding sites.
  • each EGF-like module is known to bind one Ca 2+ -ion (29-34), and the EGF-like modules have also been demonstrated to mediate Ca 2+ dependent protein/protein interaction (35).
  • a 43 kDa membrane protein ( ⁇ 2 -macroglobulin receptor-associated protein, or Heparin-binding protein) (28,36) is known to interact both with the LDL-receptor-related protein and with the rat Heymann antigen in a Ca 2+ dependent manner (28). No physiological function has yet been assigned to this protein, but it appears also in tissues where the Heymann antigen and the LDL-receptor-related proteins are not expressed (28). An interesting observation is the presence of a putative leucine-zipper motif in the aminoterminal part of the 43 kDa protein (36), considering that such motifs have been suggested to influence the opening and closure of membrane ion channels (37).
  • the 43 kDa protein interacts with the Heymann antigen, it can be assumed to form a complex also with the calcium sensor protein in a Ca 2+ dependent manner. Interaction with the 43 kDa protein might be important for the transmission of Ca 2+ induced conformational changes within the extracellular portion of the molecule to the cell interior. It is also possible that additional proteins interact with the calcium sensor in a Ca 2+ dependent manner, and that such an interaction is important for the modulation of the sensor response. The mechanisms by which an activated calcium sensor triggers further signalling to the cell interior is unknown, although we have in preliminary experiments utilized immunoprecipitation to isolate a phosphorylated form of the sensor protein in dispersed parathy roid cells loaded with [ 32 P]-ortophosphate (unpublished observation).
  • the calcium sensor protein of the placenta may be involved in maintenance of a feto-maternal Ca 2+ gradient and placental Ca 2+ transport, possibly by mediating calcium regulation of the parathyroid hormone related peptide (PTHrP) production and/or 1,25 (OH) 2 D 3 metabolism (8). Its presence already within the blastocyst (unpublished observation) may indicate a function also as adhesion molecule, or implicate involvement in differentiation or growth regulation, as suggested for the Heymann antigen (38). The function of a calcium sensor within the kidney tubule brush border is less well explored.
  • the enzyme 1- ⁇ -hydroxylase present in the placenta and proximal kidney tubule is regulated by extracellular calcium, and the calcium sensor might accordingly regulate 1,25 (OH) 2 D 3 metabolism, but it may possibly also influence Ca 2+ reabsorption from the glomerular filtrate (7-9).
  • nucleotide sequence of Fig. 3 can be used for isolating the complete DNA and gene sequence encoding the calcium sensor.
  • an analysis of overlapping cDNA clones in conjunction with PCR techniques is used.
  • the gene sequence can be obtained from the analysis of overlapping genomic cosmid and/or lambda phage clones.
  • MOLECULE TYPE protein
  • SEQUENCE DESCEIPTION SEQ ID NO:6:

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Abstract

L'invention se rapporte à l'isolation d'un clone d'ADNc codant le capteur de calcium dans le placenta humain et à l'analyse Northern blot successive confirmant l'expression de mRNA également dans la parathyroïde et dans les cellules des tubules rénaux chez l'homme. On démontre une grande similitude de séquence avec l'antigène de la néphrite de Heymann chez le rat, qui est une glycoprotéine de la bordure en brosse des tubules rénaux présentant une capacité de liaison de calcium. Les analyses immunohistochimiques montrent une distribution tissulaire de la protéine capteur de calcium similaire à celle déjà décrite pour l'antigène de Heymann. On avance la théorie que la protéine capteur de calcium identifiée constitue un capteur universel pour la reconnaissance des variations du calcium extracellulaire, et qu'elle joue un rôle clé dans la régulation du calcium par l'intermédiaire de différents systèmes corporels. Cette protéine capteur de calcium appartient à la superfamille LDL de glycoprotéines, auxquelles on attribue principalement une fonction de récepteurs protéiques, mais qui présentent également d'importantes fonctions de liaison de calcium.
PCT/SE1994/000483 1993-05-23 1994-05-24 Capteur de calcium chez l'homme WO1994028019A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
EP94917235A EP0700401A1 (fr) 1993-05-24 1994-05-24 Capteur de calcium chez l'homme
AU69021/94A AU685127B2 (en) 1993-05-24 1994-05-24 Human calcium sensor
JP6525857A JPH08510135A (ja) 1993-05-24 1994-05-24 ヒトカルシウムセンサー
US08/476,515 US6239270B1 (en) 1993-05-24 1995-06-07 Nucleic acids encoding human calcium sensor protein
US08/652,877 US6187548B1 (en) 1993-05-23 1996-05-23 Methods using human calcium sensor protein, fragments thereof and DNA encoding same

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE9301764A SE504108C2 (sv) 1993-05-24 1993-05-24 Human Kalcium sensor
SE9301764-8 1993-05-24

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US34483694A Continuation-In-Part 1993-05-23 1994-11-23

Publications (1)

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WO1994028019A1 true WO1994028019A1 (fr) 1994-12-08

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PCT/SE1994/000483 WO1994028019A1 (fr) 1993-05-23 1994-05-24 Capteur de calcium chez l'homme

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EP (1) EP0700401A1 (fr)
JP (1) JPH08510135A (fr)
AU (1) AU685127B2 (fr)
CA (1) CA2163013A1 (fr)
SE (1) SE504108C2 (fr)
WO (1) WO1994028019A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997008201A1 (fr) * 1995-08-28 1997-03-06 University Of Florida Auto-anticorps chez des patients atteints d'hypoparathyroidie acquise et methode de dosage afferente
WO1999038500A3 (fr) * 1998-01-30 2000-04-13 Brigham & Womens Hospital Recepteur humain de detection du calcium servant a detecter et a traiter le cancer
EP0792159A4 (fr) * 1994-11-23 2003-01-02 Rhone Poulenc Rorer Pharma Proteine humaine detectrice du calcium, fragments de cette proteine et adn codant pour elle

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
The Journal of Biological Chemistry, Volume 265, No. 14, May 1990, JUHLIN C. et al., "500-Kilodalton Calcium Sensor Regulating Cytoplasmic Ca2+ in Cytotrophoblast Cells of Human Placenta", page 8275 - page 8279. *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0792159A4 (fr) * 1994-11-23 2003-01-02 Rhone Poulenc Rorer Pharma Proteine humaine detectrice du calcium, fragments de cette proteine et adn codant pour elle
WO1997008201A1 (fr) * 1995-08-28 1997-03-06 University Of Florida Auto-anticorps chez des patients atteints d'hypoparathyroidie acquise et methode de dosage afferente
US5723343A (en) * 1995-08-28 1998-03-03 University Of Florida Autoantibodies in patients with acquired hypoparathyroidism and assay method therefor
US6066475A (en) * 1995-08-28 2000-05-23 University Of Florida Parathyroid cytosolic autoantigens recognized by autoantibodies from patients with acquired hypoparathyroidism
WO1999038500A3 (fr) * 1998-01-30 2000-04-13 Brigham & Womens Hospital Recepteur humain de detection du calcium servant a detecter et a traiter le cancer
US6296833B1 (en) 1998-01-30 2001-10-02 The Brigham And Women's Hospital, Inc. Human calcium-sensing receptor in the detection and treatment of cancer

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JPH08510135A (ja) 1996-10-29
EP0700401A1 (fr) 1996-03-13
SE504108C2 (sv) 1996-11-11
SE9301764D0 (sv) 1993-05-24
AU6902194A (en) 1994-12-20
SE9301764L (sv) 1994-11-25
AU685127B2 (en) 1998-01-15
CA2163013A1 (fr) 1994-12-08

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