Solubilization, purification, and partial characterization of thyrotropin receptor from bovine and human thyroid glands.
In an attempt to purify the thyroid receptor for TSH and to study interaction with the thyroid-stimulating antibody (TSAb) of graves' disease, we used both bovine and human thyroid glands. With either tissue, the 10,000 X g pellet of homogenized material was solubilized with 0.5% Triton N-101; excess Triton was removed by Amberlite XAD-2, and purification was effected by TSH affinity chromatography, followed by gel filtration on Sepharose 6B. Greatest purification was achieved with bovine tissue; this receptor preparation was 183-fold concentrated over the starting material, but contained only 6% of the original TSH-binding activity, due in part to spontaneous loss over the 4 days required for processing. On polyacrylamide gel electrophoresis, there were at least three protein bands, one of which was probably a subunit of thyroglobulin. Purified immunoglobulin G with thyroid-stimulating antibody activity inhibited the binding of TSH at all stages of purification of the receptor.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006111:Graves Disease / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D018828:Immunoglobulins, Thyroid-Stimulating; D007700:Kinetics; D011956:Receptors, Cell Surface / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D011989:Receptors, Thyrotropin; D012995:Solubility; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013961:Thyroid Gland / Q000032:analysis*; D013972:Thyrotropin / Q000378:metabolism
D000906:Antibodies; D007074:Immunoglobulin G; D018828:Immunoglobulins, Thyroid-Stimulating; D011956:Receptors, Cell Surface; D011989:Receptors, Thyrotropin; D013439:Sulfhydryl Reagents; D013972:Thyrotropin