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Title: Effect of glycoprotein-processing inhibitors on fucosylation of glycoproteins

Abstract

Influenza viral hemagglutinin contains L-fucose linked alpha 1,6 to some of the innermost GlcNAc residues of the complex oligosaccharides. To determine what structural features of the oligosaccharide were required for fucosylation influenza virus-infected MDCK cells were incubated in the presence of various inhibitors of glycoprotein processing to stop trimming at different points. After several hours of incubation with the inhibitors, (5,6-TH)fucose and (1- UC)mannose were added to label the glycoproteins, and cells were incubated in inhibitor and isotope for about 40 h to produce mature virus. Glycopeptides were prepared from the viral and the cellular glycoproteins, and these glycopeptides were isolated by gel filtration on Bio-Gel P-4. The glycopeptides were then digested with endo-beta-N-acetylglucosaminidase H and rechromatographed on the Bio-Gel column. In the presence of castanospermine or 2,5-dihydroxymethyl-3,4-dihydroxypyrrolidine, both inhibitors of glucosidase I, most of the radioactive mannose was found in Glc3Man7-9GlcNAc structures, and these did not contain radioactive fucose. In the presence of deoxymannojirimycin, an inhibitor of mannosidase I, most of the ( UC)mannose was in a Man9GlcNAc structure which was also not fucosylated. However, in the presence of swainsonine, an inhibitor of mannosidase II, the ( UC)mannose was mostly in hybrid types of oligosaccharides, and these structures alsomore » contained radioactive fucose. Treatment of the hybrid structures with endoglucosaminidase H released the (TH)fucose as a small peptide (Fuc-GlcNAc-peptide), whereas the ( UC)mannose remained with the oligosaccharide. The data support the conclusion that the addition of fucose linked alpha 1,6 to the asparagine-linked GlcNAc is dependent upon the presence of a beta 1,2-GlcNAc residue on the alpha 1,3-mannose branch of the core structure.« less

Authors:
;
Publication Date:
Research Org.:
Univ. of Texas Health Science Center, San Antonio
OSTI Identifier:
6336315
Resource Type:
Journal Article
Journal Name:
J. Biol. Chem.; (United States)
Additional Journal Information:
Journal Volume: 27
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; GLUCOPROTEINS; LABELLING; METABOLISM; HEMAGGLUTININS; MOLECULAR STRUCTURE; ALKALOIDS; CARBON 14 COMPOUNDS; CELL TRANSFORMATIONS; DOGS; ENZYME INHIBITORS; GLUCOSIDASE; HEXOSES; MANNOSE; OLIGOSACCHARIDES; TRACER TECHNIQUES; TRITIUM COMPOUNDS; AGGLUTININS; ALDEHYDES; ANIMALS; ANTIBODIES; CARBOHYDRATES; ENZYMES; GLYCOSYL HYDROLASES; HYDROLASES; ISOTOPE APPLICATIONS; LABELLED COMPOUNDS; MAMMALS; MONOSACCHARIDES; O-GLYCOSYL HYDROLASES; ORGANIC COMPOUNDS; PROTEINS; SACCHARIDES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Schwarz, P M, and Elbein, A D. Effect of glycoprotein-processing inhibitors on fucosylation of glycoproteins. United States: N. p., 1985. Web.
Schwarz, P M, & Elbein, A D. Effect of glycoprotein-processing inhibitors on fucosylation of glycoproteins. United States.
Schwarz, P M, and Elbein, A D. Mon . "Effect of glycoprotein-processing inhibitors on fucosylation of glycoproteins". United States.
@article{osti_6336315,
title = {Effect of glycoprotein-processing inhibitors on fucosylation of glycoproteins},
author = {Schwarz, P M and Elbein, A D},
abstractNote = {Influenza viral hemagglutinin contains L-fucose linked alpha 1,6 to some of the innermost GlcNAc residues of the complex oligosaccharides. To determine what structural features of the oligosaccharide were required for fucosylation influenza virus-infected MDCK cells were incubated in the presence of various inhibitors of glycoprotein processing to stop trimming at different points. After several hours of incubation with the inhibitors, (5,6-TH)fucose and (1- UC)mannose were added to label the glycoproteins, and cells were incubated in inhibitor and isotope for about 40 h to produce mature virus. Glycopeptides were prepared from the viral and the cellular glycoproteins, and these glycopeptides were isolated by gel filtration on Bio-Gel P-4. The glycopeptides were then digested with endo-beta-N-acetylglucosaminidase H and rechromatographed on the Bio-Gel column. In the presence of castanospermine or 2,5-dihydroxymethyl-3,4-dihydroxypyrrolidine, both inhibitors of glucosidase I, most of the radioactive mannose was found in Glc3Man7-9GlcNAc structures, and these did not contain radioactive fucose. In the presence of deoxymannojirimycin, an inhibitor of mannosidase I, most of the ( UC)mannose was in a Man9GlcNAc structure which was also not fucosylated. However, in the presence of swainsonine, an inhibitor of mannosidase II, the ( UC)mannose was mostly in hybrid types of oligosaccharides, and these structures also contained radioactive fucose. Treatment of the hybrid structures with endoglucosaminidase H released the (TH)fucose as a small peptide (Fuc-GlcNAc-peptide), whereas the ( UC)mannose remained with the oligosaccharide. The data support the conclusion that the addition of fucose linked alpha 1,6 to the asparagine-linked GlcNAc is dependent upon the presence of a beta 1,2-GlcNAc residue on the alpha 1,3-mannose branch of the core structure.},
doi = {},
url = {https://www.osti.gov/biblio/6336315}, journal = {J. Biol. Chem.; (United States)},
number = ,
volume = 27,
place = {United States},
year = {1985},
month = {11}
}