GLYCOSYLATION OF THE ENVELOPE GLYCOPROTEIN FROM A POLYTROPIC MURINE RETROVIRUS IN 2 DIFFERENT HOST-CELLS
GLYCOSYLATION OF THE ENVELOPE GLYCOPROTEIN FROM A POLYTROPIC MURINE RETROVIRUS IN 2 DIFFERENT HOST-CELLS
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DOI：
10.1111/j.1432-1033.1990.tb19409.x
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发表时间：
1990-11-13
期刊：
影响因子：
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通讯作者：
GEYER, R
中科院分区：
文献类型：
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作者：
GEYER, H;KEMPF, R;GEYER, R
在小鼠胚胎成纤维细胞和貂肺细胞上繁殖含有Friend貂细胞病灶诱导病毒的包膜基因加上两性鼠白血病病毒基因组其余部分的多效性重组逆转录病毒。从培养物上清液中收获用[2 - 3 H]甘露糖代谢标记的病毒颗粒，并用去污剂裂解。通过免疫亲和层析从裂解物中分离病毒包膜糖蛋白，并通过制备型SDS/PAGE纯化。通过用内切-β-N-乙酰氨基葡萄糖苷酶H和肽-N4-（N-乙酰-β-葡糖胺酰基）天冬酰胺酰胺酶F顺序处理胰蛋白酶糖肽释放寡核苷酸，并通过高效液相色谱法分级分离。结果表明，在小鼠胚胎成纤维细胞中合成的病毒糖蛋白作为主要成分携带部分岩藻糖基化的双触角、2，4-和2，具有0-4个唾液酸残基的6分支三触角和四触角复合型N-聚糖和仅少量具有5- 9个甘露糖残基。作为一个特征，部分复合型聚糖含有额外的Gal（α1-3）取代基。从貂肺细胞上繁殖的病毒体中获得的糖蛋白含有部分岩藻糖基化的双触角和2，4-分支三触角寡糖（含1-3个唾液酸残基），以及痕量的高甘露糖型物质（含8或9个甘露糖残基）。因此，结果表明，逆转录病毒包膜糖蛋白的复杂N型聚糖主要表现出细胞特异性变异，包括寡糖分支、唾液酸化和由额外Gal（α1-3）残基取代的差异。
A polytropic recombinant retrovirus containing the envelope gene of Friend mink cell focus‐inducing virus plus the remainder of the genome of an amphoropic murineleukemia virus was propagated on mouse embryo fibroblasts and mink lung cells. Virus particles, metabolically labeled with [2‐3H]mannose, were harvested from the culture supernatants and lysed with detergents. The viral envelope glycoprotein was isolated from the lysates by immunoaffinity chromatography and purified by preparative SDS/PAGE. Oligosaccharides were liberated by sequential treatment of tryptic glycopeptides with endo‐ß‐N‐acetylglucosaminidase H and peptide‐N4‐(N‐acetyl‐ß‐glucosaminyl)asparagine amidase F and fractionated by high‐performance liquid chromatography. Individual glycans were characterized chromatographically, by methylation analyses and in part, by enzymic microsequencing.The results demonstrated that viral glycoproteins, synthesized in mouse embryo fibroblasts, carried as major constituents partially fucosylated diantennary, 2,4‐ and 2,6‐branched triantennary and tetraantennary complex typeN‐glycans with 0–4 sialic acid residues and only small amounts of high‐mannose type species with 5–9 mannose residues. As a characteristic feature, part of the complex type glycans contained additional Gal(α1–3) substituents. Glycoprotein obtained from virions propagated on mink lung cells, contained partially fucosylated diantennary and 2,4‐branched triantennary oligosaccharides with 1–3 sialic acid residues, in addition to trace amounts of high‐mannose type species with 8 or 9 mannose residues. Thus, the results reveal that predominantly, the complex typeN‐glycans of the retroviral envelope glycoprotein display cell‐specific variations including differences in oligosaccharide branching, sialylation and subtitution by additional Gal(α1–3) residues.