Probing the structural states of human immunodeficiency virus type 1 Pr55gag by using monoclonal antibodies
Probing the structural states of human immunodeficiency virus type 1 Pr55gag by using monoclonal antibodies
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DOI：
10.1128/jvi.01717-07
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发表时间：
2008-03-01
影响因子：
5.4
通讯作者：
Hope, Thomas J.
中科院分区：
文献类型：
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作者：
LeBlanc, Jason J.;Perez, Omar;Hope, Thomas J.
Gag-FP(荧光蛋白)融合构建物通常用于研究人类免疫缺陷病毒1型组装，产生遍布细胞质的扩散信号和通常被描述为病毒样颗粒(VLP)的点状信号，代表已组装但未处理的Gag。然而，这些颗粒不能准确地描述为VLP，因为荧光显微镜不能提供结构分辨率。我们在这里证明，当未处理的GAG组装时，单抗p24无法与其同源表位结合，这是VLP与未组装的单体GAG的区别。此外，我们还发现，组装和未组装的断口信号沿微管传播。这些单抗研究为检测逆转录病毒组装提供了新的工具。
Gag-FP (fluorescent protein) fusion constructs are commonly used to study human immunodeficiency virus type 1 assembly, yielding diffuse signals throughout the cytoplasm along with punctate signals routinely described as virus-like particles (VLPs) representing assembled but unprocessed Gag. However, these particles cannot be accurately described as VLPs, since fluorescence microscopy cannot provide structural resolution. We demonstrate here that the inability of a monoclonal p24 antibody to bind its cognate epitope when unprocessed Gag is assembled distinguishes VLPs from unassembled, monomeric Gag. Furthermore, we show that assembled and unassembled Gag punctate signals travel along microtubules. These monoclonal antibody studies provide a new tool for examining retroviral assembly.