Catalytic activity of anti-thyroglobulin antibodies.

Thyroglobulin (Tg)-specific autoantibodies from a patient with Hashimoto's thyroiditis hydrolyzed radiolabeled Tg, shown by production of several smaller sized products on SDS electrophoresis gels. The apparent Km value for Tg was in the nanomolar range, a property typical of an Ab combining site. The Tg antibodies also hydrolyzed tripeptide-methylcoumarinamide (MCA) substrates with lower affinity, displaying a preference for Arg-MCA and Lys-MCA containing conjugates. The hydrolysis of one of these conjugates, Pro-Phe-Arg-MCA, was inhibited competitively by Tg, suggesting a catalytic site located in the Ab combining site. In control experiments, 1) the hydrolytic activities were removed by immunoadsorption with immobilized anti-human IgG; 2) IgG depleted of the Tg-specific Abs by affinity chromatography did not display Tg and Pro-Phe-Arg-MCA hydrolyzing activities; and 3) the peptide-MCA hydrolyzing activity tracked exactly with the 150-kDa IgG peak on a gel filtration column run in denaturing solvent (6 M guanidine chloride).