Prevention and reversion of protein aggregation by molecular chaperones in the E-coli cytosol:: implications for their applicability in biotechnology

被引:131
作者
Schlieker, C [1 ]
Bukau, B [1 ]
Mogk, A [1 ]
机构
[1] Inst Biochem & Mol Biol, D-79104 Freiburg, Germany
关键词
molecular chaperones; protein aggregation; inclusion bodies; DnaK; ClpB; proteases;
D O I
10.1016/S0168-1656(02)00033-0
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
The amount of a native protein reflects an equilibrium of protein synthesis, de novo folding and protein stability. Stress situations, like heat shock, or overproduction of a protein can cause an imbalance in this equilibrium, resulting in protein aggregation. Molecular chaperones control protein folding processes and protect misfolded proteins from aggregation in all cells. Since protein aggregation is frequently observed upon synthesis of heterologous proteins in E. coli, molecular chaperones have been applied in biotechnology by their co-overproduction with the desired protein. While increasing protein solubility in some cases, this approach has not been generally successful. Recent findings demonstrate, that protein aggregation, even in case of inclusion bodies, must not be a dead end in the life cycle of a protein. Such resolubilization of aggregated proteins is mediated by a bi-chaperone system consisting of CIpB and DnaK, the prokaryotic representatives of the Hsp100 and Hsp70 families. The disaggregation capacity of this bi-chaperone system has now been demonstrated in vitro and in vivo for a wide variety of aggregated proteins and offers a new perspective to increase the solubility of proteins of interest. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:13 / 21
页数:9
相关论文
共 30 条
[1]   Construction and characterization of a Helicobacter pylori clpB mutant and role of the gene in the stress response [J].
Allan, E ;
Mullany, P ;
Tabaqchali, S .
JOURNAL OF BACTERIOLOGY, 1998, 180 (02) :426-429
[2]   Dual function of protein confinement in chaperonin-assisted protein folding [J].
Brinker, A ;
Pfeifer, G ;
Kerner, MJ ;
Naylor, DJ ;
Hartl, FU ;
Hayer-Hartl, M .
CELL, 2001, 107 (02) :223-233
[3]   Getting newly synthesized proteins into shape [J].
Bukau, B ;
Deuerling, E ;
Pfund, C ;
Craig, EA .
CELL, 2000, 101 (02) :119-122
[4]   The Hsp70 and Hsp60 chaperone machines [J].
Bukau, B ;
Horwich, AL .
CELL, 1998, 92 (03) :351-366
[5]   Protein aggregation as bacterial inclusion bodies is reversible [J].
Carrió, MM ;
Villaverde, A .
FEBS LETTERS, 2001, 489 (01) :29-33
[6]   GroEL/GroES-mediated folding of a protein too large to be encapsulated [J].
Chaudhuri, TK ;
Farr, GW ;
Fenton, WA ;
Rospert, S ;
Horwich, AL .
CELL, 2001, 107 (02) :235-246
[7]   Trigger factor and DnaK cooperate in folding of newly synthesized proteins [J].
Deuerling, E ;
Schulze-Specking, A ;
Tomoyasu, T ;
Mogk, A ;
Bukau, B .
NATURE, 1999, 400 (6745) :693-696
[8]   Size-dependent disaggregation of stable protein aggregates by the DnaK chaperone machinery [J].
Diamant, S ;
Ben-Zvi, AP ;
Bukau, B ;
Goloubinoff, P .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (28) :21107-21113
[9]   Sequential mechanism of solubilization and refolding of stable protein aggregates by a bichaperone network [J].
Goloubinoff, P ;
Mogk, A ;
Ben Zvi, AP ;
Tomoyasu, T ;
Bukau, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (24) :13732-13737
[10]   Mechanisms of protein folding [J].
Grantcharova, V ;
Alm, EJ ;
Baker, D ;
Horwich, AL .
CURRENT OPINION IN STRUCTURAL BIOLOGY, 2001, 11 (01) :70-82