The interaction of radioiodinated thyrotropin with human plasma membranes from normal and diseased thyroid glands. Relation of thyrotropin binding to adenylate cyclase activity.
Plasma membranes have been purified from homogenates of normal human thyroid glands and multinodular euthyroid and Graves' goitres by discontinuous sucrose gradient centrifugation. Preparations of reasonable purity were obtained containing specific binding sites for thyrotropin and thyrotropin-sensitive adenylate cyclase. Optimum conditions for 125I-labeled thyrotropin binding were pH 7.8 and 37 degrees C. Sodium ions and concentration of Tris above 20 mM reduced thyrotropin binding. Human plasma membranes showed no species specificity toward thyrotropins from 3 different species (ox, hog and man). Displacement curves of I125-labeled bovine thyrotropin by unlabeled hormones was in the order of increasing concentrations of bovine, porcine and human highly purified thyrotropins and was inversely related to the specific biological activity of these preparations as determined by the bioassay in the mouse. Analysis of the interaction between membranes and 125I-labeled thyrotropin resulted in curvilinear Scatchard plots which can indicate the presence of two types of sites with high affinity -- low capacity (KD = 5 nM) and low affinity -- high capacity (KD = 500 nM) or site -- site interaction of the negative cooperativity type. No significant difference in binding site characteristics was found in normal and diseased glands (multinodular and Graves' goitres). A good correlation was found at equilibrium and in the conditions of adenylate cyclase assay between receptor occupancy and cyclase activation by b-thyrotropin.
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D011956:Receptors, Cell Surface; D013552:Swine; D013959:Thyroid Diseases / Q000201:enzymology / Q000378:metabolism*; D013961:Thyroid Gland / Q000201:enzymology / Q000378:metabolism*; D013972:Thyrotropin / Q000378:metabolism*
D011956:Receptors, Cell Surface; D013972:Thyrotropin; D000262:Adenylyl Cyclases