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Biotechnol Adv. 2014 May-Jun;32(3):564-74. doi: 10.1016/j.biotechadv.2014.02.003. Epub 2014 Feb 21.

Expression, stabilization and purification of membrane proteins via diverse protein synthesis systems and detergents involving cell-free associated with self-assembly peptide surfactants.

Author information

1
College of Basic Medical Sciences, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, China.
2
College of laboratory Medicine, Chongqing Medical University, Chongqing, China.
3
Sichuan Industrial Institute of Antibiotics, Chengdu University, Chengdu, China.
4
China Tobacco Gene Research Center, Zhengzhou Tobacco Research Institute, Zhengzhou, China.
5
College of Basic Medical Sciences, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing, China. Electronic address: Zhongliluo@163.com.

Abstract

G-protein coupled receptors (GPCRs) are involved in regulating most of physiological actions and metabolism in the bodies, which have become most frequently addressed therapeutic targets for various disorders and diseases. Purified GPCR-based drug discoveries have become routine that approaches to structural study, novel biophysical and biochemical function analyses. However, several bottlenecks that GPCR-directed drugs need to conquer the problems including overexpression, solubilization, and purification as well as stabilization. The breakthroughs are to obtain efficient protein yield and stabilize their functional conformation which are both urgently requiring of effective protein synthesis system methods and optimal surfactants. Cell-free protein synthesis system is superior to the high yields and post-translation modifications, and early signs of self-assembly peptide detergents also emerged to superiority in purification of membrane proteins. We herein focus several predominant protein synthesis systems and surfactants involving the novel peptide detergents, and uncover the advantages of cell-free protein synthesis system with self-assembling peptide detergents in purification of functional GPCRs. This review is useful to further study in membrane proteins as well as the new drug exploration.

KEYWORDS:

Cell-free; Extract; G-protein coupled receptors; Protein synthesis system; Self-assembly peptide surfactants

[Indexed for MEDLINE]

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