Post-expression strategies for structural investigations of membrane proteins
Post-expression strategies for structural investigations of membrane proteins
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DOI：
10.1016/j.sbi.2015.04.005
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发表时间：
2015-06-01
影响因子：
6.8
通讯作者：
Columbus, Linda
中科院分区：
文献类型：
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作者：
Columbus, Linda
目前，膜蛋白仅占蛋白质数据库的1.5%，因此，仍然是结构生物学家的一个挑战。表达、在膜模拟物（例如去污剂）中的稳定化、异质性（构象和化学）以及在膜模拟物存在下的结晶是遇到的四个主要瓶颈。作为回应，已经利用了几种表达后蛋白质修饰来促进膜蛋白的结构测定。本文综述了四种方法：有限的蛋白水解，去糖基化，半胱氨酸烷基化和赖氨酸甲基化。结合这些方法已经促进了40多个膜蛋白的结构测定，因此，是一个有用的除了膜蛋白结构生物学家的工具包。
Currently, membrane proteins only comprise 1.5% of the protein data bank and, thus, still remain a challenge for structural biologists. Expression, stabilization in membrane mimics (e.g. detergent), heterogeneity (conformational and chemical), and crystallization in the presence of a membrane mimic are four major bottlenecks encountered. In response, several post-expression protein modifications have been utilized to facilitate structure determination of membrane proteins. This review highlights four approaches: limited proteolysis, deglycosylation, cysteine alkylation, and lysine methylation. Combined these approaches have facilitated the structure determination of more than 40 membrane proteins and, therefore, are a useful addition to the membrane protein structural biologist's toolkit.