Changing single side-chains can greatly enhance the resistance of a membrane protein to irreversible inactivation

被引：48

作者：

Lau, FW

Nauli, S

Zhou, YF

Bowie, JU

机构：

[1] Univ Calif Los Angeles, Dept Chem & Biochem, Los Angeles, CA 90095 USA

[2] Univ Calif Los Angeles, Lab Struct Biol & Mol Med, Los Angeles, CA 90095 USA

来源：

JOURNAL OF MOLECULAR BIOLOGY | 1999年 / 290卷 / 02期

关键词：

diacylglycerol kinase; protein stability; cysteine scanning mutagenesis; protein engineering; protein denaturation;

D O I：

10.1006/jmbi.1999.2905

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：

The thermal inactivation rates of a set of 20 cysteine-substituted variants of the integral membrane protein diacylglycerol kinase were measured. Two Of the mutations, I53C and I70C, were found to significantly prolong the half-life of the enzyme in detergent solution. By combining the single mutants to create a double mutant, I53C/I70C, the half-life of the enzyme was improved from less than a minute at 70 degrees C to 51 minutes. These results demonstrate that individual side-chain substitutions can significantly improve the properties of membrane proteins in detergent solution. (C) 1999 Academic Press.

引用

页码：559 / 564

页数：6

共 19 条

[1] Escherichia coli diacylglycerol kinase is an evolutionarily optimized membrane enzyme and catalyzes direct phosphoryl transfer [J].