Biosynthesis of sulfated asparagine-linked complex carbohydrate units of calf thyroglobulin

Endocrinology
M J Spiro, R G Spiro

Abstract

Thyroglobulin from colloid as well as from membrane fractions became radiolabeled upon incubation of calf thyroid slices with [35S]sulfate. The identity of the sulfate-labeled molecule was established by immunoprecipitation, polyacrylamide gel electrophoresis, Bio-Gel A-5m filtration, and DEAE-cellulose chromatography. Size analysis by gel filtration of [35S]glycopeptides and hydrazine-released oligosaccharides indicated that the sulfate was primarily located in the complex (unit B) carbohydrate units of thyroglobulin. Moreover, although [35S]sulfate-labeled oligosaccharides were cleaved by N-glycanase to the same extent as those labeled with [3H]mannose, they were not released by endo-beta-N-acetylglucosaminidase under conditions that led to the complete removal of polymannose carbohydrate (unit A). The failure of 35S-labeled glycopeptides and oligosaccharides to bind to immobilized Concanavalin-A indicated that the sulfate residues in calf thyroglobulin are located in carbohydrate units with three or more branches. No evidence for the occurrence of tyrosine sulfate was found upon examination of Pronase digests of radiolabeled thyroglobulin, and chemical analyses excluded the presence of this amino acid down to a level of 0.5 ...Continue Reading

Citations

Apr 6, 1999·Comparative Biochemistry and Physiology. Part C, Pharmacology, Toxicology & Endocrinology·S G Venkatesh, V Deshpande
Feb 1, 1997·Thyroid : Official Journal of the American Thyroid Association·P ArvanS Abu Hossain
Mar 17, 2007·Critical Reviews in Toxicology·R Thomas ZoellerRochelle W Tyl
Dec 5, 1988·FEBS Letters·J P KamerlingJ F Vliegenthart
Aug 17, 1999·Biochemical and Biophysical Research Communications·M C NlendO Chabaud
Jan 27, 2010·Academic Medicine : Journal of the Association of American Medical Colleges·Leila R LaxCathy M Smith
May 27, 2005·Academic Medicine : Journal of the Association of American Medical Colleges·David A Cook

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