title: Multisite aggregation of p53 and implications for drug rescue.
abstract: Protein aggregation is involved in many diseases. Often, a unique aggregation-prone sequence polymerizes to form regular fibrils. Many oncogenic mutants of the tumor suppressor p53 rapidly aggregate but form amorphous fibrils. A peptide surrounding Ile254 is proposed to be the aggregation-driving sequence in cells. We identified several different aggregating sites from limited proteolysis of harvested aggregates and effects of mutations on kinetics and products of aggregation. We present a model whereby the amorphous nature of the aggregates results from multisite branching of polymerization after slow unfolding of the protein, which may be a common feature of aggregation of large proteins. Greatly lowering the aggregation propensity of any one single site, including the site of Ile254, by mutation did not inhibit aggregation in vitro because aggregation could still occur via the other sites. Inhibition of an individual site is, accordingly, potentially unable to prevent aggregation in vivo. However, cancer cells are specifically killed by peptides designed to inhibit the Ile254 sequence and further aggregation-driving sequences that we have found. Consistent with our proposed mechanism of aggregation, we found that such peptides did not inhibit aggregation of mutant p53 in vitro. The cytotoxicity was not eliminated by knockdown of p53 in 2D cancer cell cultures. The peptides caused rapid cell death, much faster than usually expected for p53-mediated transcription-dependent apoptosis. There may also be non-p53 targets for those peptides in cancer cells, such as p63, or the peptides may alter other interactions of partly denatured p53 with receptors.
authors.affiliation: MRC Laboratory of Molecular Biology, PNAC Division, Cambridge CB2 0QH, United Kingdom.
authors.forename: GuoZhen
authors.initials: G
authors.lastname: Wang
authors.affiliation: MRC Laboratory of Molecular Biology, PNAC Division, Cambridge CB2 0QH, United Kingdom arf25@cam.ac.uk.
authors.forename: Alan R
authors.initials: AR
authors.lastname: Fersht
mesh_terms: D006801:Humans; D008962:Models, Theoretical; D009154:Mutation; D009369:Neoplasms / Q000235:genetics; D066263:Protein Aggregation, Pathological; D000072417:Protein Domains; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000378:metabolism*
chemical_list: D016159:Tumor Suppressor Protein p53
keywords: amyloid; disease; mechanism; misfolding