Carbohydrates of human immunodeficiency virus. Structures of oligosaccharides linked to the envelope glycoprotein 120.
Human T-cells (H9), persistently infected with the HTLV-III strain of human immunodeficiency virus, were metabolically labeled with D-[2-3H]mannose or D-[6-3H]glucosamine. The viral envelope glycoprotein, gp120, was isolated either from cell lysates or from cell-free culture supernatant. After proteolytic digestion, the radiolabeled oligosaccharides were sequentially liberated from glycopeptides by treatment with endo-beta-N-acetylhexosaminidase H and peptide:N-glycosidase F. Oligosaccharides released were separated from residual (glyco)peptides and fractionated according to size, charge, and fucose content. The individual oligosaccharide species obtained were characterized by digestion with exoglycosidases and by chromatographic comparison with standard oligosaccharides. Our results demonstrate that the intracellular gp120 carries predominantly oligomannosidic glycans comprising nine or eight mannose residues. The secreted glycoprotein is equally substituted by oligomannosidic species, containing seven to nine mannose residues, and by fucosylated, partially sialylated bi- and triantennary complex-type oligosaccharides.
Geyer
H
H
Biochemisches Institut der Justus-Liebig-Universität Giessen, Federal Republic of Germany.
Holschbach
C
C
Hunsmann
G
G
Schneider
J
J
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005944:Glucosamine / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D006678:HIV / Q000378:metabolism; D015699:HIV Envelope Protein gp120; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D007163:Immunosorbent Techniques; D008358:Mannose / Q000378:metabolism; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D012191:Retroviridae Proteins / Q000032:analysis* / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism / Q000382:microbiology; D014759:Viral Envelope Proteins
D015699:HIV Envelope Protein gp120; D009844:Oligosaccharides; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D005944:Glucosamine; D008358:Mannose