Home LiteratureArticle Details
PMID: 2203739 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Interaction between heat shock protein DnaK and recombinant staphylococcal protein A.

Journal of bacteriology ·Vol. 172 ·No. 9 ·1990-09-00 ·Pages 5030-4

Hellebust H, Uhlén M, Enfors SO

Abstract

When a protein derived from the immunoglobulin G (IgG)-binding domains of staphylococcal protein A was expressed in Escherichia coli and recovered from cell extract by IgG affinity chromatography, the 69-kilodalton heat shock protein DnaK was found to be copurified. DnaK could be selectively eluted from the IgG column by ATP or by lowering the pH to 4.7. Protein A could subsequently be eluted by lowering the pH to 3.2. Thus, this procedure allows a one-step purification of both DnaK and protein A from cell extract. In vitro experiments with pure DnaK and protein A revealed that DnaK did not interfere with the IgG-binding properties of protein A but associated with its unfolded C-terminal in a salt-resistant manner. In addition, a specific interaction between DnaK and denaturated casein was found.

MeSH Terms
Binding Sites Electrophoresis, Polyacrylamide Gel Escherichia coli/genetics,metabolism Escherichia coli Proteins HSP70 Heat-Shock Proteins Heat-Shock Proteins/isolation & purification,metabolism Molecular Weight Plasmids Recombinant Proteins/isolation & purification,metabolism Staphylococcal Protein A/isolation & purification,metabolism
Chemicals
Escherichia coli Proteins HSP70 Heat-Shock Proteins Heat-Shock Proteins Recombinant Proteins Staphylococcal Protein A dnaK protein, E coli
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hellebust H
Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.
Uhlén M
Enfors S O
References (24)
24 references, click to expand
  1. The dnaK protein of Escherichia coli possesses an ATPase and autophosphorylating activity and is essential in an in vitro DNA replication system.
    Proc Natl Acad Sci U S A. 1983 Nov;80(21):6431-5 PMID: 6314326
  2. The dnaK protein modulates the heat-shock response of Escherichia coli.
    Cell. 1983 Sep;34(2):641-6 PMID: 6311435
  3. The genetics and regulation of heat-shock proteins.
    Annu Rev Genet. 1984;18:295-329 PMID: 6442118
  4. Production of abnormal proteins in E. coli stimulates transcription of lon and other heat shock genes.
    Cell. 1985 Jun;41(2):587-95 PMID: 3886165
  5. Immobilization and purification of enzymes with staphylococcal protein A gene fusion vectors.
    EMBO J. 1985 Apr;4(4):1075-80 PMID: 2990908
  6. The heat-shock response.
    Annu Rev Biochem. 1986;55:1151-91 PMID: 2427013
  7. Speculations on the functions of the major heat shock and glucose-regulated proteins.
    Cell. 1986 Sep 26;46(7):959-61 PMID: 2944601
  8. Escherichia coli contains a soluble ATP-dependent protease (Ti) distinct from protease La.
    Proc Natl Acad Sci U S A. 1987 Aug;84(16):5550-4 PMID: 3303028
  9. The grpE protein of Escherichia coli. Purification and properties.
    J Biol Chem. 1987 Dec 25;262(36):17437-42 PMID: 2826421
  10. Transient association of newly synthesized unfolded proteins with the heat-shock GroEL protein.
    Nature. 1988 Nov 17;336(6196):254-7 PMID: 2904124
  11. GroE heat-shock proteins promote assembly of foreign prokaryotic ribulose bisphosphate carboxylase oligomers in Escherichia coli.
    Nature. 1989 Jan 5;337(6202):44-7 PMID: 2562907
  12. Escherichia coli DnaK and GrpE heat shock proteins interact both in vivo and in vitro.
    J Bacteriol. 1989 Mar;171(3):1590-6 PMID: 2522091
  13. The heat-shock proteins.
    Annu Rev Genet. 1988;22:631-77 PMID: 2853609
  14. Escherichia coli heat shock gene mutants are defective in proteolysis.
    Genes Dev. 1988 Dec;2(12B):1851-8 PMID: 3149251
  15. Cellular defects caused by deletion of the Escherichia coli dnaK gene indicate roles for heat shock protein in normal metabolism.
    J Bacteriol. 1989 May;171(5):2337-46 PMID: 2651398
  16. Analysis and use of the serum albumin binding domains of streptococcal protein G.
    J Mol Recognit. 1988 Apr;1(2):69-74 PMID: 3273653
  17. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  18. Molecular basis of beta-galactosidase alpha-complementation.
    Proc Natl Acad Sci U S A. 1975 Apr;72(4):1254-7 PMID: 1093175
  19. Levels of major proteins of Escherichia coli during growth at different temperatures.
    J Bacteriol. 1979 Jul;139(1):185-94 PMID: 156716
  20. The B66.0 protein of Escherichia coli is the product of the dnaK+ gene.
    J Bacteriol. 1982 Mar;149(3):1175-7 PMID: 7037741
  21. Proteases in Escherichia coli.
    Methods Enzymol. 1981;80 Pt C:680-702 PMID: 7043205
  22. Antibodies to two major chicken heat shock proteins cross-react with similar proteins in widely divergent species.
    Mol Cell Biol. 1982 Mar;2(3):267-74 PMID: 7110134
  23. pUR 250 allows rapid chemical sequencing of both DNA strands of its inserts.
    Nucleic Acids Res. 1982 Oct 11;10(19):5765-72 PMID: 6292850
  24. Major heat shock gene of Drosophila and the Escherichia coli heat-inducible dnaK gene are homologous.
    Proc Natl Acad Sci U S A. 1984 Feb;81(3):848-52 PMID: 6322174
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.9.5030-5034.1990
Published
1990-09-00
Pages
5030-4
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC213159
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.172.9.5030-5034.1990 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2