Role of the chaperone DnaK in protein solubility and conformational quality in inclusion body-forming Escherichia coli cells

被引:40
|
作者
Martinez-Alonso, Monica
Vera, Andrea
Villaverde, Antonio [1 ]
机构
[1] Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Dept Genet & Microbiol, E-08193 Barcelona, Spain
[2] Univ Autonoma Barcelona, CIBER, Networking Ctr Bioengn Biomat & Nanomed, BBN, E-08193 Barcelona, Spain
关键词
DnaK; misfolding; aggregation; solubility; inclusion bodies;
D O I
10.1111/j.1574-6968.2007.00788.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Misfolding-prone proteins produced in bacteria usually fail to adopt their native conformation and aggregate. In cells producing folding-reluctant protein species, folding modulators are supposed to be limiting, a fact that enhances protein deposition. Therefore, coproducing DnaK or other main chaperones along with the target protein has been a common approach to gain solubility, although with very inconsistent and often discouraging results. In an attempt to understand the reason for this inconsistency, the impact of exogenous DnaK (encoded in an accompanying plasmid) on two protein features observed as indicators of protein quality, namely solubility and functionality, has been analysed here through the specific fluorescence emission of a reporter Green Fluorescent Protein (GFP). Intriguingly, GFP solubility is strongly dependent on its own yield but poorly affected by DnaK levels. On the contrary, the specific fluorescence of both soluble and insoluble GFP populations is simultaneously modulated by the availability of DnaK, with a profile that is clearly dissimilar to that shown by protein solubility. Therefore, solubility, not being coincident with the biological activity of the target protein, might not be a robust indicator of protein quality.
引用
收藏
页码:187 / 195
页数:9
相关论文
共 50 条
  • [21] Protein folding in the cytoplasm of Escherichia coli: Requirements for the DnaK-DnaJ-GrpE and GroEL-GroES molecular chaperone machines
    Thomas, JG
    Baneyx, F
    MOLECULAR MICROBIOLOGY, 1996, 21 (06) : 1185 - 1196
  • [22] Protein misfolding and inclusion body formation in recombinant Escherichia coli cells overexpressing heat-shock proteins
    Thomas, JG
    Baneyx, F
    JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (19) : 11141 - 11147
  • [23] The role of Escherichia coli type III secretion system 2 chaperone protein ygeG in pathogenesis of avian pathogenic Escherichia coli
    Yin, Lei
    Li, Qianwen
    Wang, Zeping
    Tu, Jian
    Shao, Ying
    Song, Xiangjun
    Pan, Xiaocheng
    Qi, Kezong
    RESEARCH IN VETERINARY SCIENCE, 2021, 140 : 203 - 211
  • [24] SPECIFICITY OF THE ESCHERICHIA-COLI CHAPERONE DNAK (70-KDA HEAT-SHOCK PROTEIN) FOR HYDROPHOBIC AMINO-ACIDS
    RICHARME, G
    KOHIYAMA, M
    JOURNAL OF BIOLOGICAL CHEMISTRY, 1993, 268 (32) : 24074 - 24077
  • [25] Kinetics of Inclusion Body Formation and Its Correlation with the Characteristics of Protein Aggregates in Escherichia coli
    Upadhyay, Arun K.
    Murmu, Aruna
    Singh, Anupam
    Panda, Amulya K.
    PLOS ONE, 2012, 7 (03):
  • [26] FACTORS INFLUENCING INCLUSION BODY FORMATION IN THE PRODUCTION OF A FUSED PROTEIN IN ESCHERICHIA-COLI
    STRANDBERG, L
    ENFORS, SO
    APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1991, 57 (06) : 1669 - 1674
  • [27] Refolding of G-protein alpha subunits from inclusion body expression in Escherichia coli
    McCusker, Emily Clare
    Robinson, Clifford R.
    Robinson, Anne Skaja
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 2006, 232 : 734 - 734
  • [28] Active Inclusion Bodies Protect the Target Protein from Degradation in Escherichia coli Cells
    Komolov, A. S.
    Bobrov, E. S.
    Sannikova, E. P.
    Gubaidullin, I. I.
    Chernomorova, N. O.
    Kozlov, D. G.
    APPLIED BIOCHEMISTRY AND MICROBIOLOGY, 2024, 60 (09) : 1631 - 1637
  • [29] Mutagenesis Reveals the Complex Relationships between ATPase Rate and the Chaperone Activities of Escherichia coli Heat Shock Protein 70 (Hsp70/DnaK)
    Chang, Lyra
    Thompson, Andrea D.
    Ung, Peter
    Carlson, Heather A.
    Gestwicki, Jason E.
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2010, 285 (28) : 21282 - 21291
  • [30] The effect of protein acetylation on the formation and processing of inclusion bodies and endogenous protein aggregates in Escherichia coli cells
    Kuczynska-Wisnik, Dorota
    Moruno-Algara, Maria
    Stojowska-Swedrzynska, Karolina
    Laskowska, Ewa
    MICROBIAL CELL FACTORIES, 2016, 15