title: Conversion of wild-type p53 core domain into a conformation that mimics a hot-spot mutant.
abstract: The wild-type p53 protein can be driven into a conformation corresponding to that adopted by structural mutant forms by heterodimerization with a mutant subunit. To seek partially folded states of the wild-type p53 core domain (p53C) we used high hydrostatic pressure (HP) and subzero temperatures. Aggregation of the protein was observed in parallel with its pressure denaturation at 25 and 37 degrees C. However, when HP experiments were performed at 4 degrees C, the extent of denaturation and aggregation was significantly less pronounced. On the other hand, subzero temperatures under pressure led to cold denaturation and yielded a non-aggregated, alternative conformation of p53C. Nuclear magnetic resonance (1H15N-NMR) data showed that the alternative p53C conformation resembled that of the hot-spot oncogenic mutant R248Q. This alternative state was as susceptible to denaturation and aggregation as the mutant R248Q when subjected to HP at 25 degrees C. Together these data demonstrate that wild-type p53C adopts an alternative conformation with a mutant-like stability, consistent with the dominant-negative effect caused by many mutants. This alternative conformation is likely related to inactive forms that appear in vivo, usually driven by interaction with mutant proteins. Therefore, it can be a valuable target in the search for ways to interfere with protein misfolding and hence to prevent tumor development.
authors.affiliation: Departamento de Bioquímica Médica, Centro Nacional de Ressonância Magnética Nuclear de Macromoléculas, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-590, Brazil.
authors.forename: Daniella
authors.initials: D
authors.lastname: Ishimaru
authors.forename: Lenize F
authors.initials: LF
authors.lastname: Maia
authors.forename: Larissa M
authors.initials: LM
authors.lastname: Maiolino
authors.forename: Pablo A
authors.initials: PA
authors.lastname: Quesado
authors.forename: Priscila C M
authors.initials: PC
authors.lastname: Lopez
authors.forename: Fabio C L
authors.initials: FC
authors.lastname: Almeida
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Valente
authors.forename: Jerson L
authors.initials: JL
authors.lastname: Silva
mesh_terms: D001665:Binding Sites; D002850:Chromatography, Gel; D017931:DNA Primers / Q000737:chemistry; D005453:Fluorescence; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D018716:Molecular Mimicry; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D017354:Point Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011312:Pressure; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013696:Temperature; D013816:Thermodynamics; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000235:genetics
chemical_list: D017931:DNA Primers; D016159:Tumor Suppressor Protein p53