MemStar: A one-shot Escherichia coli-based approach for high-level bacterial membrane protein production

被引：23

作者：

Lee, Chiara ^{[1
]}

Kang, Hae Joo ^{[1
]}

Hjelm, Anna ^{[2
]}

Qureshi, Abdul Aziz ^{[2
]}

Nji, Emmanuel ^{[2
]}

Choudhury, Hassanul ^{[1
,3
,4
]}

Beis, Konstantinos ^{[1
,3
,4
]}

de Gier, Jan-Willem ^{[2
]}

Drew, David ^{[1
,2
]}

机构：

[1] Univ London Imperial Coll Sci Technol & Med, Div Mol Biosci, London SW7 2AZ, England

[2] Stockholm Univ, Dept Biochem & Biophys, Ctr Biomembrane Res, SE-10691 Stockholm, Sweden

[3] Membrane Prot Lab, Diamond Light Source, Didcot OX11 0DE, Oxon, England

[4] Rutherford Appleton Lab, Didcot OX11 0DE, Oxon, England

来源：

FEBS LETTERS | 2014年 / 588卷 / 20期

基金：

瑞典研究理事会; 英国生物技术与生命科学研究理事会;

关键词：

Membrane protein production; X-ray crystallography; High-throughput; Escherichia coli; GREEN FLUORESCENT PROTEIN; CRYSTAL-STRUCTURE; MOLECULAR-MECHANISM; OVEREXPRESSION; EXPRESSION; AUTOINDUCTION; TRANSPORTER; OPTIMIZATION; PURIFICATION; STABILITY;

D O I：

10.1016/j.febslet.2014.08.025

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Optimising membrane protein production yields in Escherichia coli can be time- and resource-consuming. Here, we present a simple and effective Membrane protein Single shot amplification recipe: MemStar. This one-shot amplification recipe is based on the E. coli strain Lemo21(DE3), the PASM-5052 auto-induction medium and, contradictorily, an IPTG induction step. Using MemStar, production yields for most bacterial membrane proteins tested were improved to reach an average of 5 mg L-1 per OD600 unit, which is significantly higher than yields obtained with other common production strategies. With MemStar, we have been able to obtain new structural information for several transporters, including the sodium/proton antiporter NapA. (C) 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

引用

页码：3761 / 3769

页数：9

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