IDENTIFICATION OF AN N-LINKED GLYCAN IN THE V1-LOOP OF HIV-1 GP120 INFLUENCING NEUTRALIZATION BY ANTI-V3 ANTIBODIES AND SOLUBLE CD4
IDENTIFICATION OF AN N-LINKED GLYCAN IN THE V1-LOOP OF HIV-1 GP120 INFLUENCING NEUTRALIZATION BY ANTI-V3 ANTIBODIES AND SOLUBLE CD4
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DOI：
10.1007/bf01310789
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发表时间：
1994-01-01
影响因子：
2.7
通讯作者：
HANSEN, JES
中科院分区：
文献类型：
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作者：
GRAM, GJ;HEMMING, A;HANSEN, JES
糖基化是HIV-1 gp 120获得功能构象所必需的，并且gp 120的单个N-连接聚糖对于HIV-1在细胞培养物中的复制是重要的，但不是必需的。我们已经构建了一个突变的HIV-1感染性克隆缺乏的信号N-连接的糖基化的HIV-1 gp 120的V1环。通过SDS-凝胶电泳中突变体gp 120的迁移率增强来验证N-连接聚糖的缺乏。突变的病毒在每感染单位的gp 120含量或感染性方面均无差异，表明N-连接聚糖既不是必需的，也不影响细胞培养物中的病毒感染性。我们发现，突变的病毒缺乏N-连接的聚糖在V1环的gp 120是更耐中和单克隆抗体的V3环和中和可溶性重组CD 4（sCD 4）。两种病毒被ConA和构象依赖性人抗体IAM-2G 12同样良好地中和。这表明V1环中的N-连接聚糖调节gp 120的三维构象，而不改变分子的整体功能完整性。
Glycosylation is necessary for HIV-1 gp120 to attain a functional conformation, and individual N-linked glycans of gp120 are important, but not essential, for replication of HIV-I in cell culture. We have constructed a mutant HIV-I infectious clone lacking a signal for N-linked glycosylation in the V1-loop of HIV-1 gp120. Lack of an N-linked glycan was verified by a mobility enhancement of mutant gp120 in SDS-gel electrophoresis. The mutated virus showed no differences in either gp120 content per infectious unit or infectivity, indicating that the N-linked glycan was neither essential nor affecting viral infectivity in cell culture. We found that the mutated virus lacking an N-linked glycan in the V1-loop of gp120 was more resistant to neutralization by monoclonal antibodies to the V3-loop and neutralization by soluble recombinant CD4 (sCD4). Both viruses were equally well neutralized by ConA and a conformation dependent human antibody IAM-2G12. This suggests that the N-linked glycan in the V1-loop modulates the three-dimensional conformation of gp120, without changing the overall functional integrity of the molecule.