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Interaction of autoantibodies to thyrotropin receptor with a hydrophilic subunit of the thyrotropin receptor

Davies Jones, Eirian, Hashim, Faisal A., Kajita, Yoshihiro, Creagh, Fionuala M., Buckland, Paul Robert, Petersen, Vaughan B., Howells, Roger David and Rees Smith, Bernard 1985. Interaction of autoantibodies to thyrotropin receptor with a hydrophilic subunit of the thyrotropin receptor. Biochemical Journal 228 (1) , pp. 111-117. 10.1042/bj2280111

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Reduction of human thyroid membranes with dithiothreitol caused the release of a water-soluble glycoprotein which neutralized the thyrotropin (TSH) receptor-binding and thyroid-stimulating activities of Graves' serum. Analysis of the protein by gel filtration and sucrose density gradient centrifugation allowed estimates of 3.45 nm for the Stokes' radius, 3.6 S for the s20,w and 47 000 +/- 5000 (mean +/- S.D.; n = 4) for the Mr. The material released by dithiothreitol treatment could be crosslinked to 125I-labelled TSH coupled to N-hydroxysuccinimidyl 4-azidobenzoate (125I-HSAB-TSH), suggesting that it contained a component of the TSH receptor. Furthermore, analysis of the crosslinked material by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicated that it contained the TSH receptor A subunit (Mr 50 000). Several factors suggested therefore that the glycoprotein released by dithiothreitol treatment of human thyroid membranes was the TSH receptor A subunit. In particular, (a) both preparations were hydrophilic and were released from membranes by reduction, (b) they had similar Mr values and (c) both preparations crosslinked to 125I-HSAB-TSH. Material similar to the TSH receptor A subunit was released from thyroid membranes by treatment with papain, probably as a result of cleavage of the receptor A subunit at a site close to the interchain disulphide bridge. A similar mechanism, involving thyroid proteinases, was probably involved in release of material with similar properties to the TSH receptor A subunit during freezing and thawing of human thyroid homogenates.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
MRC Centre for Neuropsychiatric Genetics and Genomics (CNGG)
Subjects: R Medicine > R Medicine (General)
Uncontrolled Keywords: Autoantibodies/immunology, Azides, Cell Membrane/drug effects, Centrifugation, Density Gradient, Chromatography, Gel, Dithiothreitol/pharmacology, Graves Disease/blood, Humans, Papain/pharmacology, Receptors, Cell Surface/drug effects, Receptors, Cell Surface/immunology, Receptors, Thyrotropin, Thyroid Gland/drug effects
Publisher: Portland Press
ISSN: 0264-6021
Last Modified: 05 Feb 2020 03:39

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