The ATPase cycle of Hsp90 drives a molecular ‘clamp’ via transient dimerization of the N‐terminal domains
Open accessAbstract
How the ATPase activity of Heat shock protein 90 (Hsp90) is coupled to client protein activation remains obscure. Using truncation and missense mutants of Hsp90, we analysed the structural implications of its ATPase cycle. C-terminal truncation mutants lacking inherent dimerization displayed reduced ATPase activity, but dimerized in the presence of 5'-adenylamido-diphosphate (AMP-PNP), and AMP-PNP- promoted association of N-termini in intact Hsp90 dimers was demonstrated. Recruitment of p23/Sba1 to C-terminal truncation mutants also required AMP-PNP-dependent dimerization. The temperature- sensitive (ts) mutant T101I had normal ATP affinity but reduced ATPase activity and AMP-PNP-dependent N-terminal association, whereas the ts mutant T22I displayed enhanced ATPase activity and AMP-PNP-dependent N-terminal dimerization, indicating a close correlation between these properties. The locations of these residues suggest that the conformation of the 'lid' segment (residues 100-121) couples ATP binding to N-terminal association. Consistent with this, a mutation designed to favour 'lid' closure (A107N) substantially enhanced ATPase activity and N-terminal dimerization. These data show that Hsp90 has a molecular 'clamp' mechanism, similar to DNA gyrase and MutL, whose opening and closing by transient N-terminal dimerization are directly coupled to the ATPase cycle.
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A107N stabilizes NTD-MD contacts.
“we selected previously characterized mutants that either disrupt [R32A (34) and R380A (35)] or stabilize [A107N (26)] NTD-MD contacts.”
From Client binding shifts the populations of dynamic Hsp90 conformations through an allosteric network (2021) · cites this paper as [26]
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- Authors
- Prodromou, C., Panaretou, B., Chohan, S., Siligardi, G., O'Brien, R., Ladbury, J. E., Roe, S. M., Piper, P. W., & Pearl, L. H.
- Affiliations
- Institute of Cancer Research (GB)
- Imperial College London (GB)
- King's College London (GB)
- University College London (GB)
- Corresponding author
- Pearl, Laurence H.
- ORCID
- Venue
- The EMBO Journal
- Published
- 2000-08-15
- Volume
- 19
- Issue
- 16
- Pages
- 4383-4392
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC302038
- OpenAlex
- W2046044279
- Cited by
- 492
- References
- 39
- Citations by year
- 2026: 18 · 2025: 15 · 2024: 12 · 2023: 16 · 2022: 16 · 2021: 15 · 2020: 18 · 2019: 14 · 2018: 12 · 2017: 17 · 2016: 20 · 2015: 14 · 2014: 29 · 2013: 22 · 2012: 20
- FWCI
- 6.14
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- The EMBO Journal (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- UCL Discovery (University College London) (submittedVersion)
- Figshare (submittedVersion, cc-by)
- Research Portal (King's College London) (submittedVersion)
- Funders
- Wellcome Trust
- 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 › ATP Synthase and ATPases Research
- Keywords
- HSP90, ATPase cycle, molecular clamp, AMP-PNP, temperature-sensitive mutants, ATP binding, protein dimerization, DNA gyrase, MutL
- MeSH terms
- MutL Proteins, Adenosine Triphosphatases, Adenosine Triphosphate, Adenylyl Imidodiphosphate, Bacterial Proteins, Circular Dichroism, Cross-Linking Reagents, DNA Topoisomerases, Type II, Fungal Proteins, Kinetics, Models, Biological, Models, Molecular, Phenotype, Plasmids, Protein Binding, Protein Conformation, Spectrometry, Fluorescence, Temperature, Time Factors, Mutagenesis, Site-Directed, Protein Structure, Secondary, Protein Structure, Tertiary, Molecular Chaperones, HSP90 Heat-Shock Proteins, Dimerization, Mutation, Missense, DNA Gyrase, Saccharomyces cerevisiae Proteins, Escherichia coli Proteins
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