EP0493434A1 - Recepteur de thyrotropine - Google Patents

Recepteur de thyrotropine

Info

Publication number
EP0493434A1
EP0493434A1 EP90913766A EP90913766A EP0493434A1 EP 0493434 A1 EP0493434 A1 EP 0493434A1 EP 90913766 A EP90913766 A EP 90913766A EP 90913766 A EP90913766 A EP 90913766A EP 0493434 A1 EP0493434 A1 EP 0493434A1
Authority
EP
European Patent Office
Prior art keywords
tsh
tsh receptor
receptor
mammalian
protein
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP90913766A
Other languages
German (de)
English (en)
Other versions
EP0493434A4 (en
Inventor
Roger Cone
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
New England Medical Center Hospitals Inc
Original Assignee
New England Medical Center Hospitals Inc
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 New England Medical Center Hospitals Inc filed Critical New England Medical Center Hospitals Inc
Publication of EP0493434A1 publication Critical patent/EP0493434A1/fr
Publication of EP0493434A4 publication Critical patent/EP0493434A4/en
Withdrawn legal-status Critical Current

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
    • C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
    • C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor

Definitions

  • This invention concerns nucleic acid encoding a mammalian thyroid stimulating hormone (TSH, also known as thyrotropin) receptor, and purified mammalian TSH receptors.
  • TSH thyroid stimulating hormone
  • the TSH receptor is a protein believed to be involved in a human autoimmune disease termed "Graves*" disease. It is believed that antibodies against the TSH receptor are made in patients suffering from this disease. These auto-antibodies are currently detected by providing radiolabeled TSH, and detecting blocking of binding of the TSH to crude porcine membranes thought to include a TSH receptor.
  • the invention features purified nucleic acid encoding a protein having the immunological or biological activity of a mammalian TSH receptor.
  • the purified nucleic acid can be purified cDNA, or a purified vector including that nucleic acid.
  • the invention features purified, e.g., recombinant, protein having the immunological or biological activity of a mammalian TSH receptor.
  • nucleic acid or protein is provided separated from contaminating nucleic acid or other cell components, such as proteins and carbohydrates, with which the naturally occurring nucleic acid encoding the receptor occurs.
  • nucleic acid is provided as a homogeneous solution separated from all cell components, or is the major nucleic acid present in a preparation. More preferably, the nucleic acid is provided within a vector which is resident within a cell in a manner which allows expression of the nucleic acid to provide sufficient TSH receptor to be useful in this invention.
  • recombinant is meant that the protein is expressed from nucleic acid which has been manipulated by recombinant DNA methodology to place it in a vector or chromosome at a location in which it does not naturally occur.
  • the purified protein is present at a purity of at least 10% of the "total protein in a preparation, or even at 50% or 90% purity.
  • the biological activity of mammalian TSH receptor is that activity naturally associated with the TSH receptor of mammals, i.e., the ability of that protein to recognize and interact with TSH. It preferably includes other biological activities of the TSH receptor such as activating adenylate cyclase, well known to those of ordinary skill in the art.
  • the TSH receptor is that receptor occurring in humans; the nucleic acid has a nucleotide sequence encoding an amino acid sequence identical to that of a naturally occurring mammalian TSH receptor, most preferably a human TSH receptor; or the nucleic acid encodes a protein having only conservative amino acid substitutions compared to a naturally occurring mammalian TSH receptor.
  • conservative amino acid substitutions are well known to those skilled in the art and would include, for example, substitution of valine for glycine or leucine, substitution of a positively charged amino acid for another positively charged amino acid, or substitution of a negatively charged amino acid for another negatively charged amino acid.
  • substitutions will not significantly affect the biological activity of the encoded TSH receptor; i.e., the biological activity of the substituted form will be at least 75% that of the naturally occurring form.
  • the proteins of the invention can be used in a method for detecting the presence of anti-TSH receptor antibodies in the serum of a patient.
  • the method includes providing a purified TSH receptor as described above, and contacting that receptor with the serum. Reaction of the receptor with the serum is an indication of the presence of anti-TSH antibodies in that serum.
  • This method may include any of many well known immunological procedures for detection of antibodies, such as ELISA, Western blot or competitive binding assays.
  • the present invention provides a sufficient amount of a mammalian TSH receptor to be useful for rapid testing of patients for the presence of anti-TSH receptor antibodies. It also provides sufficient receptor protein to allow analysis of the sequence of the protein. Such analysis will aid determination of specific epitopes on that protein to allow design of small homologous peptides which will block the activity of autoimmune antibodies. Those peptides will thus block overstimulation of the thyroid in patients, such as those suffering from Graves' disease.
  • the invention also provides the tools necessary to allow development of agonists or antagonists of TSH binding to a mammalian TSH receptor. These antagonists will be useful for preventing hyperthyroidism due to elevated levels of TSH.
  • the invention features a method for determining the presence of TSH in a sample.
  • the method includes providing a mammalian cell having DNA encoding biologically active TSH receptor, the cell expressing TSH receptor from the DNA under assay conditions; contacting the cell with the sample to cause TSH within the sample to contact the cell; and measuring the level of intracellular cyclic adenosine monophosphate prior to and after the contacting step. An elevated level of cyclic adenosine monophosphate after the contacting step compared to prior to the contacting step is indicative of the presence of TSH within the cell.
  • Fig. 1 is a depiction of the nucleotide base sequence of the rat LH receptor probe.
  • Fig. 2 is a depiction of the nucleotide base sequence of the human LH receptor cDNA and the derived amino acid sequence.
  • Fig. 3 is a diagrammatic representation of the structure of the human LH receptor-encoding gene and the derived amino acid sequence.
  • Fig. 4 is a depiction of the nucleotide base sequence of degenerate oligonucleotide probes based on the human LH receptor DNA sequence.
  • Fig. 5 is a depiction of the partial nucleotide base sequence of the bovine and human TSH receptors. The boxed sequences indicate regions with possible sequence errors due to compression during sequence determination.
  • Fig. 6 is a depiction of the nucleotide base sequence and the derived amino acid sequence of the human TSH receptor cDNA.
  • Fig. 7 illustrates a darkfield photomicrograph (75x magnification) showing an autoradiographic signal (bright spots) produced by radiolabeled anti-sense transcript of human TSH receptor overlying a haematoxylin and eosin stained section of human thyroid.
  • Fig. 8 is a diagrammatic representation of the pATH3-hTSHR expression vector.
  • Fig. 9 illustrates a photograph of a polyacrylamide gel demonstrating the expression of the trp E-TSH receptor fusion protein (small arrow) in the absence (-) and presence (+) of indoleacetic acid.
  • the small arrow indicates a protein of the size predicted for the fusion protein.
  • Fig. 10 is a graphical representation of the the level of intracellular cyclic adenosine monophosphate (cAMP) as a function of the concentration of applied hormone.
  • TSH Receptor TSH receptors useful in this invention include any such receptor isolated from a mammal, or any protein having the biological activity of such a receptor. Such proteins will include proteins derived from naturally occurring TSH receptors having one or more of their amino acids modified conservatively as discussed above.
  • Such modification may be by any standard procedure, for example, by recombinant DNA technology.
  • receptors will be expressed by recombinant DNA technology by isolating the gene encoding that receptor, and placing that gene within an expression vector, after removing any intronic DNA that may be deleterious to expression of the full length receptor protein.
  • expression vectors will include bacterial, fungal, insect, and mammalian expression vectors which may be expressed within a bacterial, fungal, insect, or mammalian cell by techniques well known to those with ordinary skill in the art.
  • Purified mammalian TSH receptor may be also be isolated by preparing antibodies to one of the above recombinant mammalian TSH receptors, and using those antibodies to immunoaffinity purify a naturally occurring TSH receptor. Generally, such a procedure is not preferred, since the yield of TSH receptor will be extremely small.
  • proteins having the biological activity of the receptor may be designed by standard procedure. For example, oligonucleotides may be synthesized by standard procedure, and inserted into any standard expression vector to cause expression of fragments of the naturally occurring TSH receptor. These fragments can be screened by standard procedure to determine whether they have the desired biological activity of the receptor protein.
  • TSH receptor-encoding gene it may be determined by affinity chromatography, Western blot analysis, or some equivalent analysis, whether that synthetic peptide is able to bind with antibodies against a TSH receptor. Those fragments which can bind are useful in this invention.
  • the expressed TSH receptor, or that purified as described above may be fragmented by use of enzymes, e.g., trypsin, which specifically cleaves the amino acid sequence into smaller fragments. These fragments may then be tested in much the same way as the synthetic peptide fragments to determine their usefulness in methods of this invention.
  • enzymes e.g., trypsin
  • a 622 nucleotide fragment of the rat luteinizing hormone (LH) receptor gene was obtained from Deborah Segaloff of the Center for Biological Research at the Population Step, New York, New York, 10021, and from Peter Seeburg at the University of Heidelberg.
  • This DNA fragment was used as a probe of a lambda-gtll cDNA library constructed from RNA isolated from the thyroid of a patient suffering from Graves' disease. The nucleotide base sequence of this probe is shown in Fig. 1.
  • the cDNA library was constructed generally as follows. RNA of the thyroid was isolated using a standard guanidiu /thiocyanate procedure and reverse transcribed using the method of Gubler and Hoffman.
  • the resulting cDNA was size selected using a Sepharose G50 gel filtration column to select cDNA of greater than 1 kb in size.
  • the cDNA was methylated with EcoRI methylase, linked to EcoRI linkers, and then treated with EcoRI.
  • the resulting DNA was ligated to EcoRI treated lambda- gtll DNA.
  • the resulting lambda DNA was amplified in E. coli strain 1090.
  • the rat LH gene fragment was labeled with 32 P- dCTP and plates containing the lambda gtll library screened on nitrocellulose filters at low stringency in 30% formamide, 1M NaCl, at 42°C. The filters were then washed at low stringency in 2 x SSC at 50°C.
  • the strongly reacting plaques were purified three times using standard procedure, and four were determined to encode overlapping parts of the same gene by restriction endonuclease mapping, and DNA sequencing procedures.
  • the 5' terminal 600 nucleotides of the gene showed high homology to the rat LH receptor.
  • Further analysis determined that the cDNA encoded the full length human LH receptor protein with several introns remaining.
  • the nucleotide base sequence is provided in Fig. 2.
  • the amino acid sequence, molecular weight and isoelectric point of the encoded protein can be calculated by standard techniques from this sequence.
  • the encoded protein has 90% homology in amino acid sequence to the rat LH receptor protein.
  • the cDNA includes intronic DNA.
  • RNA protection experiments, Northern analysis, and polymerase chain reaction experiments showed that the mRNA encoded by this clone -is expressed in the thyroid, testes, and ovary, as well as in Graves* thyroid, and in thyroid cell lines.
  • the RNA is expressed in the thyroid but is incompletely spliced. Thus, this clone does not encode thyroid specific DNA.
  • two degenerate oligonucleotide probes were constructed, one having homology to the transmembrane domain III of the above cloned human LH receptor DNA and the other having homology to the transmembrane domain VI of the LH receptor DNA.
  • oligonucleotides were synthesized and purified by standard procedure; their sequences are shown in Fig. 4.
  • This reaction was treated at 94 ⁇ C for one minute in the presence of Taq DNA polymerase and then two minutes at 50°C and three minutes at 72°C. This cycle of heating and cooling between 50°C and 94°C was repeated thirty times. At this point, no amplification product could be observed. Five ⁇ l of the resulting reaction was removed and the procedure repeated. At this point, a DNA product was observed. No such product was observed in reactions using total RNA isolated from osteosarcoma, testes, ovary, melanoma, or placenta. Thus, the DNA product appears to be thyroid specific. The resulting material was precipitated and resuspended by standard procedure, and digested with HindiII and EcoRI.
  • the EcoRI HindiII fragment was subcloned into the vector pBS ⁇ (Strategene, La Jolla, CA) , and transformed into E. coli.
  • the resulting vector was sequenced by Sanger dideoxy procedures.
  • Both human and bovine cDNAs were sequenced and found to encode a protein having about 84% homology. Their tentative sequences are presented in Fig. 5. In contrast, the DNA had only about 68% homology with rat, porcine, and human LH receptor.
  • the fragments derived from the polymerase chain reaction were removed from the vector and labeled with 32 P. These fragments were then used as probes to screen the above described lambda-gtll library at high stringency.
  • the conditions were 50% formamide at 42°C in the presence of 1 M NaCl for 15-20 hours, and then washing of the nitrocellulose filters at 20-25°C in 2 x SSC for 15, minutes at 68°C in 1 x SSC for 45 minutes, and at 68°C in 0.1 x SSC for 45 minutes. Strongly hybridizing plaques were detected at a higher frequency than had been detected for the LH receptor clones. Twelve of these plaques were purified three times, purified DNA isolated from six, and analyzed by EcoRI restriction analysis.
  • This RNA has a 3'-untranslated sequence of between 2 and 2.5 kb, and a 5'-untranslated sequence of approximately 50 bases.
  • TR.12.6- 1 has been determined, by DNA sequencing, to contain a full length human TSH receptor cDNA (Fig. 6) .
  • the above-described cDNA from human and bovine, or any other mammal may be expressed by standard procedures to provide large quantities of TSH receptor.
  • the above cDNA may be inserted into a trp E- fusion plasmid, e.g., pATH-1, 2, or 3, to form a stable hybrid protein with the Trp E protein.
  • the cDNA may be inserted into a mammalian expression system such as a cytomegalovirus or retrovirus vector. Glycosylated protein will result when the DNA is expressed in the mammalian expression system.
  • coli transformed with pATH3- hTSHR were grown in selective M9 media for 2 hours in either the absence or presence of 40 ⁇ g/ ⁇ l indoleacetic acid, an inducer of the Trp E gene.
  • Bacterial pellets were lysed in SDS loading buffer and 1/lOth of the material was electrophoresed on a 10% polyacrylamide Laemmeli gel (Fig. 9) .
  • nucleic acid encoding TSH receptor may be used to express large quantities of TSH receptor.
  • high level expression is achieved with a Baculovirus vector pVL941, the E_ s _ coli vector paTH3, and the mammalian vector pLJ.
  • Such protein is useful for detection of auto-antibodies found in Graves' patients. This allows determination of the state of the thyroid of those patients, and indicates the progress of that patient.
  • This test may be performed in an ELISA format, for example, in a dipstick assay. The test might also take the form of a competitive binding assay employing radiolabeled TSH and TSH receptor. Such assays are extremely sensitive, and more readily performed than prior methods of detecting such antibodies.
  • the expressed protein is useful for defining the epitopes recognized by antibodies in Graves* patients. This analysis may be performed by standard procedure, for example, by expressing portions of the cloned DNA to provide partial TSH receptor fragments, or by fragmenting the expressed receptor protein as discussed above. Once the region recognized by such antibodies is defined, these fragments may be used in immunoassay procedures. In addition, definition of epitopes may be performed by. manipulating the cloned genes using standard techniques of molecular biology to provide proteins in which one or more amino acids which may form a part of one or more epitopes of the protein is altered or deleted.
  • the protein or portions thereof is also useful as a therapeutic where it may be administered in a pharmaceutically acceptable compound at a sufficient dose to alleviate one or more symptoms of Grave's patients, or other patients suffering from thyroid malfunction. Generally, such administration will be at a level between one and one thousand micrograms per kilogram of patient.
  • Small peptides may be designed which will block the activity of auto-antibodies that act as TSH agonists, and thus block stimulation of the thyroid. Other small peptides may be designed which will block auto-antibodies that act as TSH antagonists. In addition, antagonists of TSH may be constructed which prevent binding of TSH to the TSH receptor and thus prevent elevated thyroid activity. Assays for TSH
  • the first assay technique is based upon the expression of TSH receptor within a cell which does not naturally contains such a receptor. This cell, when contacted with TSH, will increase expression of cyclic adenosine monophosphate, which can be detected as a measure of the amount of TSH in a sample.
  • the human TSH receptor-encoding DNA is inserted with a mammalian retroviral vector pLJ at the BamHI to Sail sites.
  • the resulting vector is then transfected into human 2 ⁇ *3 cells and clonal cell lines containing the vector isolated by selection in the presence of the antibiotic G418.
  • Such transfection causes the cells to become responsive to TSH as measured by the activation of adenylate cyclase and accumulation of cAMP following treatment with TSH.
  • these cell lines provide a highly sensitive assay system for the hormone TSH.
  • Cells in culture or cell membrane preparations may be exposed to the sample thought to contain TSH and the resulting adenylate cyclase activity quantitated and correlated with the cyclase activity from standard dilution curves of TSH in order to calculate the concentration of TSH in a sample. Concentrations as low as 1 ng/ml or even 0.1 ng/ml can be detected in this assay.
  • This assay demonstrates that the TSH receptor encoded by the cDNA described above is biologically active and leads to specific TSH responsiveness in a previously unresponsive cell line. These cell lines are responsive not only to naturally occurring TSH but also to recombinant TSH.
  • a retrovirus expression vector pLJ (Korman et al., Proc. Natl. Acad. Sci. USA 84:2150, 1987) containing the entire tr.12 cDNA sequence was transfected into human 293 cells and intracellular cAMP concentrations measured 60 hours later using a 3 H-cAMP displacement assay after treatment with hCG, hFSH, or hTSH.
  • hFSH or hCG has little effect while the same amount of hTSH elevated intracellular cAMP over 6-fold.
  • Half maximal intracellular concentrations of cAMP were obtained with approximately 60 picomolar hTSH.
  • a 15-fold elevation of intracellular cAMP was induced by application of 100 ng/ml hTSH.
  • Transfection of the retrovirus vector alone, with no hTSH-r insert produced no elevation of intracellular cAMP over background in cells treated with 100 ng/ml TSH.
  • Expression of the human LH/CG receptor was attempted using identical methods, however, no elevation of cAMP was seen after treatment with any of the glycoprotein hormones. This could result from any of number of problems, including, for example, the deletion found in clone tr.13, or perhaps inefficient removal of the LH/CG-R introns in the non-gonadal 293 cell line.
  • the TSH receptor of this invention can be used to measure TSH by means of a competitive binding assay.
  • TSH receptor or a portion thereof capable of binding TSH, is immobilized on a support matrix.
  • the immobilized receptor is incubated with excess TSH, which has been tagged with a radioactive or florescent label, long enough for the binding reaction to come to equilibrium. Unbound TSH is removed by a washing step, and the receptor is incubated with the test sample. Once this second binding step has come to equilibrium, the immobilized receptor is washed again.
  • the amount of tagged TSH displace by TSH in the test sample then serves as a measure of the TSH present in the test sample.
  • Applicant's assignee represents that the ATCC is a depository affording permanence of the deposit and ready accessibility thereto by the public if a patent is granted. All restrictions on the availability to the public of the material so deposited will be irrevocably removed upon the granting of a patent. The material will be available during the pendency of the patent application to one determined by the Commissioner to be entitled thereto under 37 CFR 1.14 and 35 USC 122.
  • the deposited material will be maintained with all the care necessary to keep it viable and uncontaminated for a period of at least five years after the most recent request for the furnishing of a sample of the deposited microorganism, and in any case, for a period of at least thirty (30) years after the date of deposit or for the enforceable life of the patent, whichever period is longer.
  • Applicants' assignee acknowledges its duty to replace the deposit should the depository be unable to furnish a sample when requested due to the condition of the deposit.
  • Other embodiments are within the following claims

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

Abstract

Protéine présentant l'activité biologique d'un récepteur de thyrotropine mammifère, et acide nucléique purifié codant ladite protéine.
EP19900913766 1989-09-08 1990-09-07 Tsh receptor Withdrawn EP0493434A4 (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US40489989A 1989-09-08 1989-09-08
US404899 1989-09-08
US56566990A 1990-08-10 1990-08-10
US565669 1990-08-10

Publications (2)

Publication Number Publication Date
EP0493434A1 true EP0493434A1 (fr) 1992-07-08
EP0493434A4 EP0493434A4 (en) 1993-05-05

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Family Applications (1)

Application Number Title Priority Date Filing Date
EP19900913766 Withdrawn EP0493434A4 (en) 1989-09-08 1990-09-07 Tsh receptor

Country Status (4)

Country Link
EP (1) EP0493434A4 (fr)
JP (1) JP3181582B2 (fr)
CA (1) CA2066263C (fr)
WO (1) WO1991003483A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5614363A (en) * 1990-01-25 1997-03-25 New England Medical Center Hospitals, Inc. TSH receptor
US5744348A (en) * 1989-09-08 1998-04-28 New England Medical Center Hospitals, Inc. TSH receptor

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04505103A (ja) * 1989-05-05 1992-09-10 ジェネンテク，インコーポレイテッド 糖タンパク質ホルモン受容体分子
US6261800B1 (en) 1989-05-05 2001-07-17 Genentech, Inc. Luteinizing hormone/choriogonadotropin (LH/CG) receptor
ES2092998T3 (es) * 1989-12-14 1996-12-16 Brahms Diagnostica Gmbh Polipeptidos que tienen actividad de receptores de tirotropina, secuencias de acidos nucleicos que codifican para tales receptores y polipeptidos, y aplicaciones de estos polipeptidos.
EP0506890A4 (en) * 1989-12-20 1993-06-30 Nichols Institute Diagnostics Recombinant thyrotropin receptor
DE19645729C1 (de) * 1996-11-06 1998-06-04 Brahms Diagnostica Gmbh Rezeptorbindungsassay, für den Rezeptorbindungsassay geeigneter rekombinanter Fusionsrezeptor, Vektor zu dessen Herstellung sowie Reagenziensatz für die Durchführung des Rezeptorbindungsassays
DE19651093C2 (de) * 1996-12-09 1999-06-10 Brahms Diagnostica Gmbh Rezeptorbindungsassay zum Nachweis von TSH-Rezeptor-Autoantikörpern sowie Reagenziensatz für die Durchführung eines solchen Rezeptorbindungsassays
WO1999015545A1 (fr) * 1997-09-24 1999-04-01 Merck & Co., Inc. Recepteur hormonal glycoproteique aomf05 couple par proteines g
GB9823397D0 (en) 1998-10-27 1998-12-23 Rsr Ltd Assays for thyroid autoantibodies
WO2004048415A1 (fr) 2002-11-26 2004-06-10 B.R.A.H.M.S Aktiengesellschaft Mise en evidence d'auto-anticorps recepteurs tsh avec des anticorps sans affinite

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990013643A2 (fr) * 1989-05-05 1990-11-15 Genentech, Inc. Molecules receptrices d'hormone de glycoproteine
WO1991009121A2 (fr) * 1989-12-14 1991-06-27 Henning Berlin Gmbh Chemie- Und Pharmawerk Polypeptides possedant de recepteurs actifs de thyrotropine, sequences d'acide nucleique codant pour ces recepteurs et polypetides, et applications de ces polypeptides
WO1991009137A1 (fr) * 1989-12-20 1991-06-27 Basil Rapoport Recepteur de thyrotropine recombinant
WO1991010735A2 (fr) * 1990-01-15 1991-07-25 Institut National De La Sante Et De La Recherche Medicale Recepteur humain de la tsh. sequence codant pour ce recepteur

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990013643A2 (fr) * 1989-05-05 1990-11-15 Genentech, Inc. Molecules receptrices d'hormone de glycoproteine
WO1991009121A2 (fr) * 1989-12-14 1991-06-27 Henning Berlin Gmbh Chemie- Und Pharmawerk Polypeptides possedant de recepteurs actifs de thyrotropine, sequences d'acide nucleique codant pour ces recepteurs et polypetides, et applications de ces polypeptides
WO1991009137A1 (fr) * 1989-12-20 1991-06-27 Basil Rapoport Recepteur de thyrotropine recombinant
WO1991010735A2 (fr) * 1990-01-15 1991-07-25 Institut National De La Sante Et De La Recherche Medicale Recepteur humain de la tsh. sequence codant pour ce recepteur

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
64TH MEETING OF THE AMERICAN THYROID ASSOCIATION, 1989, abstract no. T-51; L. FRAZIER-SEABROOK et al.: "Isolation of a thyroid-specific cDNA with over 80% aminoacid homology with the luteinizing hormone receptor" *
ACTA ENDOCRINOLOGICA, vol. 115, suppl. 281, 1987, pages 166-172, Copenhagen, DK; J. CHAN et al.: "TSH receptor structure" *
BIOCHEM. BIOPHYS. RES. COMMUN., vol. 165, no. 3, 29th December 1989, pages 1184-1190, Academic Press, New York, US; Y. NAGAYAMA et al.: "Molecular cloning, sequence and functional expression of the cDNA for the human thyrotropin receptor" *
BIOCHEM. BIOPHYS. RES. COMMUN., vol. 165, no. 3, 29th December 1989, pages 1250-1255, Academic Press, New York, US; F. LIBERT et al.: "Cloning, sequencing and expression of the human thyrotropin (TSH) receptor: evidence for binding of autoantibodies" *
CLINICAL RESEARCH, vol. 29, no. 2, April 1981, page 294A, Thorofare, NJ, US; M.N. ISLAM et al.: "Purification of the human thyrotropin receptor" *
ENDOCRINOLOGY, vol. 110, no. 4, April 1982, pages 1381-1391, The Endocrine Soc., Baltimore, US; Y. KOIZUMI et al.: "Solubilisation, purification, and partial characterization of thyrotropin receptor from bovine and human thyroid glands" *
SCIENCE, vol. 246, 22nd December 1989, pages 1620-1622, AAAS, Washington, DC, US; M. PARMENTIER et al.: "Molecular cloning of the thyroid receptor" *
See also references of WO9103483A1 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5744348A (en) * 1989-09-08 1998-04-28 New England Medical Center Hospitals, Inc. TSH receptor
US5614363A (en) * 1990-01-25 1997-03-25 New England Medical Center Hospitals, Inc. TSH receptor

Also Published As

Publication number Publication date
JPH05501952A (ja) 1993-04-15
CA2066263C (fr) 1999-11-02
WO1991003483A1 (fr) 1991-03-21
EP0493434A4 (en) 1993-05-05
JP3181582B2 (ja) 2001-07-03
CA2066263A1 (fr) 1991-03-09

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