{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T20:50:51Z","timestamp":1761511851882},"reference-count":31,"publisher":"The Endocrine Society","issue":"1","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2004,1,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>A recent report of major pathophysiological significance, and opposed to present concepts, is that TSH (but not MS-1, a hamster monoclonal thyroid-stimulating antibody), cleaves the single-chain TSH receptor (TSHR) on the cell surface into its two-subunit form. We reassessed the issue using two approaches. First we wished to confirm the flow-cytometric assay previously used to quantitate TSHR cleavage. We used CHO cell lines expressing large (TSHR-10,000 cells) or conventional (TSHR-0 cells) numbers of TSHR. Cells were preincubated (16 h) in either control medium or medium supplemented with TSH (5 \u00d7 10\u22128m) or MS-1 (10 \u03bcg\/ml). After stringent washing to maximize removal of residual ligand, we performed flow cytometry with two antibodies, one recognizing only the single-chain TSHR, the other recognizing all (cleaved and uncleaved) TSHRs. TSH pretreatment did not appear to increase TSHR cleavage. Instead we observed ligand occupancy of the TSHR (with MS-1) or fewer receptors on the cell surface (down-regulation), particularly with the TSHR-0 cells. Second, we covalently cross-linked [125I]TSH to monolayers of these cells, an unequivocal method to determine directly the proportion of single-chain and two-subunit TSHR forms. Pretreatment of TSHR-10,000 and TSHR-0 cells with TSH had no effect on the degree of TSHR cleavage. MS-1 slightly reduced spontaneous cleavage. In conclusion, in contrast to a recent report, we show that TSH does not alter the subunit structure of its cognate receptor, and we provide insight into the difficulties associated with the flow-cytometric assay for TSHR cleavage.<\/jats:p>","DOI":"10.1210\/en.2003-1002","type":"journal-article","created":{"date-parts":[[2003,9,2]],"date-time":"2003-09-02T00:41:59Z","timestamp":1062463319000},"page":"4-10","source":"Crossref","is-referenced-by-count":12,"title":["Does Thyrotropin Cleave Its Cognate Receptor?"],"prefix":"10.1210","volume":"145","author":[{"given":"Gregorio D.","family":"Chazenbalk","sequence":"first","affiliation":[{"name":"Autoimmune Disease Unit, Cedars-Sinai Research Institute and School of Medicine, University of California, Los Angeles, Los Angeles, California 90048"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chun-Rong","family":"Chen","sequence":"additional","affiliation":[{"name":"Autoimmune Disease Unit, Cedars-Sinai Research Institute and School of Medicine, University of California, Los Angeles, Los Angeles, California 90048"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sandra M.","family":"McLachlan","sequence":"additional","affiliation":[{"name":"Autoimmune Disease Unit, Cedars-Sinai Research Institute and School of Medicine, University of California, Los Angeles, Los Angeles, California 90048"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Basil","family":"Rapoport","sequence":"additional","affiliation":[{"name":"Autoimmune Disease Unit, Cedars-Sinai Research Institute and School of Medicine, University of California, Los Angeles, Los Angeles, California 90048"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"80","reference":[{"key":"2020071621284786200_R1","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/0014-5793(82)80176-2","article-title":"Photo-affinity labelling of the thyrotropin 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Fluorescent protein-tagged hRSHR reveals post-translational complexes.","volume":"276","author":"Latif","year":"2001","journal-title":"J Biol Chem"},{"key":"2020071621284786200_R12","doi-asserted-by":"crossref","first-page":"4516","DOI":"10.1210\/endo.140.10.7031","article-title":"On the functional importance of thyrotropin receptor intramolecular cleavage.","volume":"140","author":"Chazenbalk","year":"1999","journal-title":"Endocrinology"},{"key":"2020071621284786200_R13","doi-asserted-by":"crossref","first-page":"1324","DOI":"10.1210\/en.2002-0005","article-title":"Targeted restoration of cleavage in a noncleaving thyrotropin receptor demonstrates that cleavage is insufficient to enhance ligand-independent activity.","volume":"144","author":"Chen","year":"2003","journal-title":"Endocrinology"},{"key":"2020071621284786200_R14","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1111\/j.1432-1033.1995.0338k.x","article-title":"In Chinese hamster ovary K1 cells dog and human 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