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

Journal Article · · Endocrinology; (United States)
OSTI ID:6954445
;
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 residues/polypeptide subunit. Studies with density gradient-separated membrane fractions as well as with puromycin indicated that sulfate addition is a late event in thyroglobulin biosynthesis which occurs in the Golgi compartment. Furthermore, it was observed that the nondimerized thyroglobulin subunit was much less sulfate labeled than the mature molecule.
Research Organization:
Harvard Medical School, Boston, MA (USA)
OSTI ID:
6954445
Journal Information:
Endocrinology; (United States), Journal Name: Endocrinology; (United States) Vol. 123:1; ISSN ENDOA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AGGLUTININS
ALDEHYDES
AMIDES
AMINO ACIDS
ANIMALS
ANTIBODIES
ASPARAGINE
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
CALVES
CARBOHYDRATES
CARBOXYLIC ACIDS
CATTLE
CHROMATOGRAPHY
CONCANAVALIN
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
ELECTROPHORESIS
ENZYMES
EVEN-ODD NUCLEI
GLOBULINS
GLUCOSE
GLYCOSYL HYDROLASES
HEMAGGLUTININS
HEXOSES
HYDROLASES
IN VITRO
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LECTINS
LEUCINE
LIGHT NUCLEI
MAMMALS
MANNOSE
MONOSACCHARIDES
NUCLEI
OLIGOSACCHARIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXYGEN COMPOUNDS
PEPTIDE HYDROLASES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RUMINANTS
SACCHARIDES
SEPARATION PROCESSES
SERINE PROTEINASES
SULFATES
SULFUR 35
SULFUR COMPOUNDS
SULFUR ISOTOPES
SYNTHESIS
THYROGLOBULIN
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TRYPSIN
VERTEBRATES