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Interaction of E. coli Hsp90 with DnaK Involves the DnaJ Binding Region of DnaK.
- Source :
-
Journal of molecular biology [J Mol Biol] 2017 Mar 24; Vol. 429 (6), pp. 858-872. Date of Electronic Publication: 2016 Dec 21. - Publication Year :
- 2017
-
Abstract
- The 90-kDa heat shock protein (Hsp90) is a widely conserved and ubiquitous molecular chaperone that participates in ATP-dependent protein remodeling in both eukaryotes and prokaryotes. It functions in conjunction with Hsp70 and the Hsp70 cochaperones, an Hsp40 (J-protein) and a nucleotide exchange factor. In Escherichia coli, the functional collaboration between Hsp90 <subscript>Ec</subscript> and Hsp70, DnaK, requires that the two chaperones directly interact. We used molecular docking to model the interaction of Hsp90 <subscript>Ec</subscript> and DnaK. The top-ranked docked model predicted that a region in the nucleotide-binding domain (NBD) of DnaK interacted with a region in the middle domain of Hsp90 <subscript>Ec</subscript> . We then made substitution mutants in DnaK residues suggested by the model to interact with Hsp90 <subscript>Ec</subscript> . Of the 12 mutants tested, 11 were defective or partially defective in their ability to interact with Hsp90 <subscript>Ec</subscript> in vivo in a bacterial two-hybrid assay and in vitro in a bio-layer interferometry assay. These DnaK mutants were also defective in their ability to function collaboratively in protein remodeling with Hsp90 <subscript>Ec</subscript> but retained the ability to act with DnaK cochaperones. Taken together, these results suggest that a specific region in the NBD of DnaK is involved in the interaction with Hsp90 <subscript>Ec</subscript> , and this interaction is functionally important. Moreover, the region of DnaK that we found to be necessary for Hsp90 <subscript>Ec</subscript> binding includes residues that are also involved in J-protein binding, suggesting a functional interplay among DnaK, DnaK cochaperones, and Hsp90 <subscript>Ec</subscript> .<br /> (Published by Elsevier Ltd.)
- Subjects :
- DNA Mutational Analysis
Escherichia coli genetics
Escherichia coli Proteins genetics
HSP70 Heat-Shock Proteins genetics
Molecular Docking Simulation
Protein Binding
Two-Hybrid System Techniques
Escherichia coli enzymology
Escherichia coli Proteins metabolism
HSP70 Heat-Shock Proteins metabolism
HSP90 Heat-Shock Proteins metabolism
Protein Interaction Mapping
Subjects
Details
- Language :
- English
- ISSN :
- 1089-8638
- Volume :
- 429
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Journal of molecular biology
- Publication Type :
- Academic Journal
- Accession number :
- 28013030
- Full Text :
- https://doi.org/10.1016/j.jmb.2016.12.014