Purification and characterization of HIV-human protein complexes
Purification and characterization of HIV-human protein complexes
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DOI：
10.1016/j.ymeth.2010.08.007
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发表时间：
2011-01-01
期刊：
影响因子：
4.8
通讯作者：
Krogan, Nevan J.
中科院分区：
文献类型：
--
作者：
Jaeger, Stefanie;Gulbahce, Natali;Krogan, Nevan J.
为了充分了解病原体如何感染宿主和劫持关键生物学过程，系统地绘制病原体内部和病原体-宿主蛋白质-蛋白质相互作用(PPI)的图谱是至关重要的。由于病毒基因组相对较小(通常在10-100个蛋白质左右)，可以使用不同的实验平台生成全面的宿主病毒PPI图谱，包括亲和标签纯化-质谱仪(AP-MS)和酵母双杂交(Y2H)方法。这样的全球图谱为病毒进入、复制和组装的分子机制提供了公正的见解。然而，到目前为止，只有两种杂交方法被系统地用于表征病毒-宿主蛋白-蛋白相互作用，尽管从文献中手动挑选单个宿主-病原体PPI的数据库中存在大量数据。我们将总结这项工作，并描述一个AP-MS平台，该平台可用于表征病毒-人类蛋白复合体，并讨论其在HIV基因组中的应用。(C)2010年，由爱思唯尔公司出版。
To fully understand how pathogens infect their host and hijack key biological processes, systematic mapping of intra-pathogenic and pathogen-host protein-protein interactions (PPIs) is crucial. Due to the relatively small size of viral genomes (usually around 10-100 proteins), generation of comprehensive host-virus PPI maps using different experimental platforms, including affinity tag purification-mass spectrometry (AP-MS) and yeast two-hybrid (Y2H) approaches, can be achieved. Global maps such as these provide unbiased insight into the molecular mechanisms of viral entry, replication and assembly. However, to date, only two-hybrid methodology has been used in a systematic fashion to characterize viral-host protein-protein interactions, although a deluge of data exists in databases that manually curate from the literature individual host-pathogen PPIs. We will summarize this work and also describe an AP-MS platform that can be used to characterize viral-human protein complexes and discuss its application for the HIV genome. (c) 2010 Published by Elsevier Inc.