Lack of complex N-glycans on HIV-1 envelope glycoproteins preserves protein conformation and entry function
Lack of complex N-glycans on HIV-1 envelope glycoproteins preserves protein conformation and entry function
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DOI：
10.1016/j.virol.2010.02.019
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发表时间：
2010-06-05
期刊：
影响因子：
3.7
通讯作者：
Sanders, Rogier W.
中科院分区：
文献类型：
--
作者：
Eggink, Dirk;Melchers, Mark;Sanders, Rogier W.
HIV-1包膜糖蛋白复合物（Env）是旨在引发体液免疫的疫苗开发的焦点。Env的广泛和异质的N-连接糖基化影响折叠、与凝集素受体的结合、抗原性和免疫原性。我们的特点是重组Env蛋白和病毒颗粒在哺乳动物细胞中产生的，缺乏N-乙酰葡糖胺转移酶I（GnTI），一种酶的寡甘露糖N-聚糖的复杂N-聚糖的转化所必需的。碳水化合物分析显示，在GnTI（-/-）细胞中产生的三聚体Env仅含有寡甘露糖N-聚糖，主要是不完全修剪的寡甘露糖聚糖。糖基化修饰对Env蛋白的折叠和构象影响不大。在GnTI（-/-）细胞中产生的病毒具有感染性，表明转化为复合聚糖对于Env进入功能不是必需的，尽管病毒与C型凝集素DC-SIGN的结合增强。操纵Env的N-糖基化可用于结构和功能研究以及疫苗设计。(C)2010年爱思唯尔公司All rights reserved.
The HIV-1 envelope glycoprotein complex (Env) is the focus of vaccine development aimed at eliciting humoral immunity. Env's extensive and heterogeneous N-linked glycosylation affects folding, binding to lectin receptors, antigenicity and immunogenicity. We characterized recombinant Env proteins and virus particles produced in mammalian cells that lack N-acetylglucosaminyltransferase I (GnTI), an enzyme necessary for the conversion of oligomannose N-glycans to complex N-glycans. Carbohydrate analyses revealed that trimeric Env produced in GnTI(-/-) cells contained exclusively oligomannose N-glycans, with incompletely trimmed oligomannose glycans predominating. The folding and conformation of Env proteins was little affected by the manipulation of the glycosylation. Viruses produced in GnTI(-/-) cells were infectious, indicating that the conversion to complex glycans is not necessary for Env entry function, although virus binding to the C-type lectin DC-SIGN was enhanced. Manipulating Env's N-glycosylation may be useful for structural and functional studies and for vaccine design. (C) 2010 Elsevier Inc. All rights reserved.