Identification of Conserved Residues Required for the Binding of a Tetratricopeptide Repeat Domain to Heat Shock Protein 90
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The sequential binding of heat shock protein 90 (hsp90) to a series of tetratricopeptide repeat (TPR) proteins is critical to its function as a molecular chaperone. We have used site-directed mutagenesis to clarify the structural basis for the binding of hsp90 to the TPR domain of phosphoprotein phosphatase 5 (PP5). This TPR domain was chosen for study because its three-dimensional structure is known. We examined co-immunoprecipitation of hsp90 with wild type and mutant TPR constructs from transfected cells. Only mutations located on one face of the TPR domain affected hsp90 binding. This allowed the identification of a binding groove. Three basic residues that are highly conserved in hsp90-binding TPR proteins extend prominently into this groove. Lys-97 and Arg-101 were absolutely required for hsp90 binding, while mutation of Arg-74 diminished, but did not abrogate, hsp90 binding. Mutation of Lys-32, another conserved basic residue in the binding groove, also blocked hsp90 binding. The TPR domain of PP5 bound specifically to a 12-kDa C-terminal fragment of hsp90. This binding was reduced by mutation of acidic residues in the hsp90 fragment. These data suggest conservation, among hsp90-binding TPR proteins, of a binding groove containing basic residues that interact with acidic residues near the C terminus of hsp90.
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Mutations in the TPR domain (M2) disrupt FKBP51 binding to Hsp90.
“Mutations in the TPR domain that disrupt binding to Hsp90, described originally by Chinkers and associates in PP5 (29), were also constructed (M2).”
From FKBP51 Promotes Assembly of the Hsp90 Chaperone Complex and Regulates Androgen Receptor Signaling in Prostate Cancer Cells (2010) · cites this paper as [29]
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- Authors
- Russell, L. C., Whitt, S. R., Chen, M.-S., & Chinkers, M.
- Affiliations
- University of South Alabama (US)
- Corresponding author
- Chinkers, Michael
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1999-07
- Volume
- 274
- Issue
- 29
- Pages
- 20060-20063
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1983211885
- Cited by
- 165
- References
- 36
- Citations by year
- 2026: 3 · 2025: 2 · 2024: 2 · 2023: 6 · 2022: 3 · 2021: 7 · 2020: 2 · 2019: 2 · 2018: 3 · 2017: 3 · 2015: 3 · 2014: 4 · 2013: 6 · 2012: 7
- FWCI
- 3.34
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01hl047063-08)
- National Heart, Lung, and Blood Institute (HL47063)
- National Institute of Diabetes and Digestive and Kidney Diseases (DK 55877, R01DK055877)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- HSP90, tetratricopeptide repeat domain, protein-protein binding, binding groove, acidic residues, molecular chaperones, site-directed mutagenesis
- MeSH terms
- Amino Acid Sequence, Animals, Cytoplasm, HeLa Cells, Humans, Peptides, Phosphoprotein Phosphatases, Protein Binding, Mutagenesis, Site-Directed, Conserved Sequence, HSP90 Heat-Shock Proteins, COS Cells
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