The impact of dnaKJ overexpression on recombinant protein solubility results from antagonistic effects on the control of protein quality

被引：10

作者：

Petersson, L ^{[1
]}

Carrió, MM ^{[1
]}

Vera, A ^{[1
]}

Villaverde, A ^{[1
]}

机构：

[1] Univ Autonoma Barcelona, Dept Genet & Microbiol, Inst Biotecnol & Biomed, E-08193 Barcelona, Spain

来源：

BIOTECHNOLOGY LETTERS | 2004年 / 26卷 / 07期

关键词：

chaperones; DnaK; E; coli; GroEL; inclusion bodies; protein aggregation;

D O I：

10.1023/B:BILE.0000021963.31863.a4

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We have produced increasing levels of DnaK and its co-chaperone DnaJ along with the model VP1LAC misfolding-prone protein, to explore the role of DnaK on the management of Escherichia coli inclusion bodies. While relative solubility of VP1LAC is progressively enhanced, the heat-shock response is down-regulated as revealed by decreasing levels of GroEL. This is accompanied by an increasing yield of VP1LAC and a non-regular evolution of its insoluble fraction, at moderate levels of DnaK resulting in more abundant inclusion bodies. Also, the impact of chaperone co-expression is much more pronounced in wild type cells than in a DnaK(-) mutant, probably due to the different background of heat shock proteins in these cells. The involvement of DnaK in the supervision of misfolding proteins is then pictured as a dynamic balance between its immediate holding and folding activities, and the side-effect downregulation of the heat shock response though the limitation of other chaperone and proteases activities.

引用

页码：595 / 601

页数：7

共 26 条

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