Solution structure of p53 core domain: Structural basis for its instability
Open accessAbstract
The 25-kDa core domain of the tumor suppressor p53 is inherently unstable and melts at just above body temperature, which makes it susceptible to oncogenic mutations that inactivate it by lowering its stability. We determined its structure in solution using state-of-the-art isotopic labeling techniques and NMR spectroscopy to complement its crystal structure. The structure was very similar to that in the crystal but far more mobile than expected. Importantly, we were able to analyze by NMR the structural environment of several buried polar groups, which indicated structural reasons for the instability. NMR spectroscopy, with its ability to detect protons, located buried hydroxyl and sulfhydryl groups that form suboptimal hydrogen-bond networks. We mutated one such buried pair, Tyr-236 and Thr-253 to Phe-236 and Ile-253 (as found in the paralogs p63 and p73), and stabilized p53 by 1.6 kcal/mol. We also detected differences in the conformation of a mobile loop that might reflect the existence of physiologically relevant alternative conformations. The effects of temperature on the dynamics of aromatic residues indicated that the protein also experiences several dynamic processes that might be related to the presence of alternative hydrogen-bond patterns in the protein interior. p53 appears to have evolved to be dynamic and unstable.
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The p53-DBD is in equilibrium with partially unfolded structures, indicative of a thermodynamically unstable protein.
“In addition, its CD spectrum reveals a high content of random coil, indicating that the p53-DBD is in equilibrium with partially unfolded structures, indicative of a thermodynamically unstable protein (Fig. 4, B and C) (25).”
From Client binding shifts the populations of dynamic Hsp90 conformations through an allosteric network (2021) · cites this paper as [25]
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Client binding shifts the populations of dynamic Hsp90 conformations through an allosteric network
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- Authors
- Cañadillas, J. M. P., Tidow, H., Freund, S. M. V., Rutherford, T. J., Ang, H. C., & Fersht, A. R.
- Affiliations
- Medical Research Council Centre for Protein Engineering, Medical Research Council Centre, Hills Road, Cambridge CB2 2QH, United Kingdom
- Medical Research Council (GB)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2006-02-06
- Volume
- 103
- Issue
- 7
- Pages
- 2109-2114
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC1413739
- OpenAlex
- W2109998141
- Cited by
- 174
- References
- 26
- Citations by year
- 2026: 6 · 2025: 6 · 2024: 6 · 2023: 6 · 2022: 15 · 2021: 11 · 2020: 7 · 2019: 10 · 2018: 7 · 2017: 7 · 2016: 11 · 2015: 6 · 2014: 5 · 2013: 8 · 2012: 7
- FWCI
- 5.30
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Cancer Research UK
- Medical Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Physical Sciences › Chemistry › Spectroscopy › Advanced NMR Techniques and Applications
- Keywords
- p53 core domain, protein instability, solution structure, hydrogen bond network, protein stabilization, alternative conformations, tumor suppressor
- MeSH terms
- Crystallography, Humans, Mutation, Magnetic Resonance Spectroscopy, Solutions, Thermodynamics, Water, Tumor Suppressor Protein p53, Protein Structure, Tertiary, Hydrophobic and Hydrophilic Interactions
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