Structural Distortion of p53 by the Mutation R249S and its Rescue by a Designed Peptide: Implications for "mutant Conformation"

Assaf Friedler, Brian S. DeDecker, Stefan M.V. Freund, Caroline Blair, Stefan Rüdiger, Alan R. Fersht*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

64 Scopus citations

Abstract

Missense mutations in the DNA-binding core domain of the tumour suppressor protein p53 are frequent in cancer. Many of them result in loss of native structure. The mutation R249S is one of the six most common cancer-associated p53 mutations ("hot-spots"). As it is highly frequent in hepatocellular carcinoma, its rescue is an important therapeutic target. We have used NMR techniques to study the structural effects of the R249S mutation. The overall fold of the core domain is retained in R249S, and it does not take up a denatured "mutant conformation". However, the β-sandwich had increased flexibility and, according to changes in chemical shift, there was local distortion throughout the DNA-binding interface. It is likely that the R249S mutation resulted in an ensemble of native and native-like conformations in a dynamic equilibrium. The peptide FL-CDB3 that was designed to rescue mutants of p53 by binding specifically to its native structure was found to revert the chemical shifts of R249S back towards the wild-type values and so reverse the structural effects of mutation. We discuss the implications for a rescue strategy and also for the analysis of antibody-binding data.

Original languageAmerican English
Pages (from-to)187-196
Number of pages10
JournalJournal of Molecular Biology
Volume336
Issue number1
DOIs
StatePublished - 6 Feb 2004
Externally publishedYes

Bibliographical note

Funding Information:
This work was supported by Cancer Research UK and the MRC. A.F. was supported by a long-term fellowship (no. LT00056/2000-M) from the Human Frontier Science Program. We thank Mark Bycroft for helpful discussions and advice and Karoly von Glos for peptide synthesis. S.R. was supported by a Marie Curie Fellowship from the EU.

Keywords

  • Cancer
  • Mutant
  • NMR
  • Peptide
  • p53

Fingerprint

Dive into the research topics of 'Structural Distortion of p53 by the Mutation R249S and its Rescue by a Designed Peptide: Implications for "mutant Conformation"'. Together they form a unique fingerprint.

Cite this