gp160 of HIV-I synthesized by persistently infected Molt-3 cells is terminally glycosylated: evidence that cleavage of gp160 occurs subsequent to oligosaccharide processing.
The envelope glycoprotein of HIV-I in infected, cultured human T cells is synthesized as a precursor of apparent Mr 160 kDa (gp160) and is cleaved to two glycoproteins, gp120 and gp41, which are the mature envelope glycoproteins in the virus. Neither the temporal and spatial features of glycosylation nor the oligosaccharide processing and proteolytic cleavage of the envelope glycoprotein are well understood. To understand more about these events, we investigated the glycosylation and cleavage of the envelope glycoproteins in the CD4+ human cell line, Molt-3, persistently infected with HIV-I (HTLV IIIB). The carbohydrate analysis of gp160 and gp120 and the behavior of the glycoproteins and glycopeptides derived from them on immobilized lectins demonstrate that both of these glycoproteins contain complex- and high-mannose-type Asn-linked oligosaccharides. In addition, the N-glycanase-resistant oligosaccharides of gp120 were found to contain N-acetyl-galactosamine, a common constituent of Ser/Thr-linked oligosaccharides. Pulse-chase analysis of the conversion of [35S]cysteine-labeled gp160 showed that in Molt-3 cells it takes about 2 h for gp120 to arise with a half-time of conversion of about 5 h. At its earliest detectable occurrence, gp120 was found to contain complex-type Asn-linked oligosaccharides. Taken together, these results indicate that proteolytic cleavage of gp160 to gp120 and gp41 occurs either within the trans-Golgi or in a distal compartment.
Merkle
R K
RK
Department of Biochemistry, University of Georgia, Athens 30602.
Helland
D E
DE
Welles
J L
JL
Shilatifard
A
A
Haseltine
W A
WA
Cummings
R D
RD
D001216:Asparagine / Q000378:metabolism; D002460:Cell Line; D005690:Galactose / Q000378:metabolism; D015686:Gene Products, env / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D008358:Mannose / Q000378:metabolism; D019158:N-Acetylneuraminic Acid; D009844:Oligosaccharides / Q000378:metabolism; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012794:Sialic Acids / Q000378:metabolism
D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D009844:Oligosaccharides; D011498:Protein Precursors; D012794:Sialic Acids; D001216:Asparagine; D019158:N-Acetylneuraminic Acid; D008358:Mannose; D005690:Galactose