Isolation of desialylated and low-iodinated thyroglobulin in an experimental rat thyroid tumour.

Abstract

ABSTRACT In an experimental rat thyroid tumour (Wollman line 1-1C2) thyroglobulin (TG) was isolated from soluble iodoproteins by ammonium sulphate fractionation and density gradient ultracentrifugation. The isolated TG was characterized by polyacrylamide gel electrophoresis, microimmunodiffusion, immunoelectrophoresis, analytical ultracentrifugation and by chemical determination of sialic acid and iodine content. Tumour thyroglobulin has a sedimentation coefficient of 17.7S, which is similar to that of enzymatically desialylated thyroglobulin (18.4S) in the normal rat; tumour TG shows a slower electrophoretic mobility than native TG and similar to rat desialylated TG. In double immunodiffusion tumour TG is shown to precipitate with an identity reaction with both native and desialylated rat TG; in immunoelectrophoresis it has a slower migration than native TG and is very close to that of enzymatically desialylated TG. Tumour TG has an iodine content of 0.02 % and a sialic content of 0.15 %. Chromatography of the 125I-labelled tumour TG revealed the presence of mono- and diiodothyrosines, a very small amount of triiodothyronine but no thyroxine. These results suggest that tumour TG, isolated in the experimental rat thyroid tumour, previously shown not to incorporate sialic acid into thyroglobulin, has a different electrophoretic mobility from native TG but is very similar to desialylated rat TG and has a very low sialic acid and iodine content as well as a striking decrease in thyroxine.