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. 1990 Sep;172(9):5030–5034. doi: 10.1128/jb.172.9.5030-5034.1990

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

H Hellebust 1, M Uhlén 1, S O Enfors 1
PMCID: PMC213159  PMID: 2203739

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.

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