Cloning, expression, purification, and characterization of the membrane protein UncI from Escherichia coli

被引：3
|
作者：
Hartmann, Claudia [2 ]
Engel, Andreas [1 ,2 ]
机构：
[1] Case Western Reserve Univ, Dept Pharmacol, Cleveland, OH 44106 USA
[2] Univ Basel, Biozentrum, Ctr Cellular Imaging & Nano Analyt, CH-4058 Basel, Switzerland
来源：
PROTEIN EXPRESSION AND PURIFICATION | 2011年 / 79卷 / 02期
基金：
瑞士国家科学基金会;
关键词：
Membrane protein expression; UncI; ATP synthase; Detergent screen; Gel filtration; ADENOSINE-TRIPHOSPHATASE COMPLEX; ATP-SYNTHASE; NUCLEOTIDE-SEQUENCE; 1ST GENE; OPERON; SUBUNIT; PRODUCT;
D O I：
10.1016/j.pep.2011.05.017
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
The Escherichia coli unc-operon encodes the genes for the subunits of the F0F1-ATP synthase and an integral membrane protein of unknown function called UncI. UncI influences the cell-growth and activity of F0F1, but its exact function is still unknown. The expression level is too low to extract milligram amounts of UncI from E. coli membranes and the existing purification protocol based on methanol/chloroform is not suitable for structural and functional studies. Here we present protocols to increase the expression level, to purify UncI in a detergent where UncI is monodisperse, and we characterize its oligomeric state. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码：187 / 190
页数：4
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