Structural characterization of the N-linked oligosaccharides derived from HIVgp120 expressed in lepidopteran cells

被引:12
作者
Butters, TD
Yudkin, B
Jacob, GS
Jones, IM
机构
[1] Univ Oxford, Glycobiol Inst, Dept Biochem, Oxford OX1 3QU, England
[2] Univ Oxford, Dept Biochem, Oxford OX1 3QU, England
[3] Monsanto Co, St Louis, MO USA
[4] NERC, Inst Virol, Oxford OX1 3SR, England
关键词
fucosylated glycans; recombinant glycoprotein; insect cells;
D O I
10.1023/A:1006999718552
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The oligosaccharides of recombinant HIV gp120 expressed in lepidopteran Sf9 cells were analysed after hydrazine release by gel permeation and high pH anion exchange chromatography. N-Linked glycans were exclusively of the oligomannose series and no evidence for charged complex or hybrid type glycans was found. However a glycosylation reaction similar to those found in vertebrates was evident. The major glycoform of gp120, that comprised 30% of all the species analysed, was structurally identified by exoglycosidase digestion and found to be a core fucosylated structure, Man alpha 1,6(Man alpha 1,3)Man beta 1,4GlcNAc(Fuc alpha 1,6)GlcNAc. Further confirmation of the ability of lepidopteran cells to fucosylate N-linked glycans was provided by an in vitro analysis of this reaction using authentic oligosaccharide substrates.
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页码:83 / 88
页数:6
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