Current pivotal strategies leading a difficult target protein to a sample suitable for crystallographic analysis
Current pivotal strategies leading a difficult target protein to a sample suitable for crystallographic analysis
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目前的关键策略是将困难的目标蛋白转化为适合晶体分析的样品
DOI：
10.1042/bst20200106
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Yamashita A
中科院分区：
文献类型：
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作者：
原 幸大;橋本 博;和田 啓;岡本麻里，杉浦勝明，小熊圭祐，猪熊 壽，遠藤麻衣子，THI Le Dung，山下(川西)奈那子，芳賀 猛;Yamashita A
晶体结构分析是确定蛋白质结构的基本方法。然而，目标蛋白质的结晶是分析中最困难的过程。该过程在样品制备（包括表达和纯化）期间经常受到阻碍。即使在样品被纯化后，也不是所有的候选蛋白质都结晶。在这篇小型综述中，对目前用于克服蛋白质结晶过程中遇到的障碍的方法进行了分类。具体而言，将有效结晶的策略与管道进行比较，其中通过预结晶筛选评估各种表达宿主和构建体、纯化和结晶条件以及作为靶特异性结合蛋白的结晶分子伴侣。这些方法也在不断发展，以改进这一进程。所述方法可用于晶体学分析和其他结构测定技术（例如低温电子显微镜）中的样品制备。
Crystallographic structural analysis is an essential method for the determination of protein structure. However, crystallization of a protein of interest is the most difficult process in the analysis. The process is often hampered during the sample preparation, including expression and purification. Even after a sample has been purified, not all candidate proteins crystallize. In this mini-review, the current methodologies used to overcome obstacles encountered during protein crystallization are sorted. Specifically, the strategy for an effective crystallization is compared with a pipeline where various expression hosts and constructs, purification and crystallization conditions, and crystallization chaperones as target-specific binder proteins are assessed by a precrystallization screening. These methodologies are also developed continuously to improve the process. The described methods are useful for sample preparation in crystallographic analysis and other structure determination techniques, such as cryo-electron microscopy.