title: Structural distortion of p53 by the mutation R249S and its rescue by a designed peptide: implications for "mutant conformation".
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 beta-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.
authors.affiliation: Cambridge University Chemical Laboratory and Cambridge Centre for Protein Engineering, MRC Centre, Hills Road, Cambridge CB2 2QH, UK.
authors.forename: Assaf
authors.initials: A
authors.lastname: Friedler
authors.forename: Brian S
authors.initials: BS
authors.lastname: DeDecker
authors.forename: Stefan M V
authors.initials: SM
authors.lastname: Freund
authors.forename: Caroline
authors.initials: C
authors.lastname: Blair
authors.forename: Stefan
authors.initials: S
authors.lastname: Rüdiger
authors.forename: Alan R
authors.initials: AR
authors.lastname: Fersht
mesh_terms: D001665:Binding Sites; D006801:Humans; D008958:Models, Molecular; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D017354:Point Mutation; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: D010455:Peptides; D016159:Tumor Suppressor Protein p53