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Topography and polypeptide distribution of terminal N-acetylglucosamine residues on the surfaces of intact cells. Evidence for O-linked GlcNAc

Thesis/Dissertation ·
OSTI ID:7117845
Bovine milk galactosyltransferase has been used, in conjunction with UDP-({sup 3}H) galactose, as an impermeant probe for accessible GlcNAc residues on the surfaces of lymphocytes, red blood cells and macrophages. Kinetics of labeling are biphasic, leveling off at approximately 30 minutes. The data strongly indicate vectorial surface labeling and covalent attachment of galactose. Thymocytes, T-lymphocytes, and B-lymphocytes have approximately 10{sup 6}, 3 {times} 10{sup 6}, and 5 {times} 10{sup 6} galactosylatable sites on their cell surfaces, respectively. Numerous proteins are exo-galactosylated that differ quantitatively among the major functional subsets of lymphocytes. In thymocytes 49% of the exo-galactosylated oligosaccharides are alkali-labile, whereas 80% and 90% of that derived from T-lymphocytes and B-lymphocytes can be {beta}-eliminated, respectively. Composition, size, and HPLC on two types of high-resolution columns establish that the bulk of the exo-galactosylated, {beta}-eliminated oligosaccharides are Gal{beta}1-4GlcNAcitol. These data suggest the presence of O-glycosidically linked GlcNAc monosaccharide on many lymphocyte cell-surface proteins. These studies not only describe dramatic differences in the amounts and distribution of terminal GlcNAc residues on phenotypically different lymphocyte populations, but they also describe the presence of a novel protein-saccharide linkage, which is present on numerous lymphocyte proteins.
Research Organization:
Johns Hopkins Univ., Baltimore, MD (USA)
OSTI ID:
7117845
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
AMINES
AMINO ACIDS
ANIMAL CELLS
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOHYDRATES
CARBOXYLIC ACIDS
CHROMATOGRAPHY
CONNECTIVE TISSUE CELLS
DISTRIBUTION
ENZYMES
ERYTHROCYTES
GALACTOSE
GLUCOSAMINE
GLYCINE
GLYCOSYL TRANSFERASES
HEXOSAMINES
HEXOSES
HEXOSYL TRANSFERASES
HYDROGEN COMPOUNDS
KINETICS
LABELLING
LEUKOCYTES
LIQUID COLUMN CHROMATOGRAPHY
LYMPHOCYTES
MACROPHAGES
MATERIALS
MONOSACCHARIDES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PEPTIDES
PHAGOCYTES
PHENOTYPE
POLYPEPTIDES
PROTEINS
REACTION KINETICS
SACCHARIDES
SEPARATION PROCESSES
SOMATIC CELLS
SUBCELLULAR DISTRIBUTION
THYMOCYTES
TRANSFERASES
TRITIUM COMPOUNDS