Identification of novel interactions in HIV-1 capsid protein assembly by high-resolution mass spectrometry
Identification of novel interactions in HIV-1 capsid protein assembly by high-resolution mass spectrometry
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DOI：
10.1016/s0022-2836(02)01245-7
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发表时间：
2003-01-24
影响因子：
5.6
通讯作者：
Prevelige, PE
中科院分区：
文献类型：
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作者：
Lanman, J;Lam, TT;Prevelige, PE
未成熟和成熟HIV-1病毒粒子的多形性使得使用传统方法难以表征亚基间的相互作用。虽然已知孤立结构域的结构，但挑战在于确定亚基间的相互作用，从而将这些结构域打包成超分子结构。采用高分辨率质谱法测定了可溶性衣壳蛋白（CA）和体外组装的CA的酰胺氢交换保护因子。通过对保护系数的比较和化学交联实验，绘制出了组装管中亚基/亚基界面图。这一分析为从CA管的冷冻电镜图像重建中提出的同型N域和C域相互作用提供了直接的生化证据。最重要的是，我们发现了一个以前未被认识到的亚基间。N域- c域相互作用。这种相互作用的检测调和了以前不一致的生物物理和遗传数据。2003爱思唯尔科学有限公司版权所有。
The pleomorphic nature of the immature and mature HIV-1 virions has made it difficult to characterize intersubunit interactions using traditional approaches. While the structures of isolated domains are known, the challenge is to identify intersubunit interactions and thereby pack these domains into supramolecular structures. Using high-resolution mass spectrometry, we have measured the amide hydrogen exchange protection factors for the soluble capsid protein (CA) and CA assembled in vitro. Comparison of the protection factors as well as chemical crosslinking experiments has led to a map of the subunit/subunit interfaces in the assembled tubes. This analysis provides direct biochemical evidence for the homotypic N domain and C domain interactions proposed from cryo-electron microscopy image reconstruction of CA tubes. Most significantly, we have identified a previously unrecognized intersubunit. N domain-C domain interaction. The detection of this interaction reconciles previously discrepant biophysical and genetic data. (C) 2003 Elsevier Science Ltd. All rights reserved.