title: Conformational changes in p53 analysed using new antibodies to the core DNA binding domain of the protein.
abstract: The p53 protein contains a protease resistant core section that binds to DNA in a sequence specific manner and whose crystal structure has been determined. This core is flanked at the N-terminus by the transcriptional transactivation domain and at the C-terminus by sequences involved in the oligomerisation of the protein. Extensive immunochemical analysis of p53 has shown that dominant antigenic sites lie within these N- and C-terminal domains while few antibodies to the central core have been identified. One of these, PAb240, has been extensively characterised as its epitope is cryptic in the native DNA binding core structure but is exposed by denaturation. This epitope is also exposed on many p53 proteins that contain point mutations in the core domain suggesting that these mutations may have a common affect on the structure of the core. To investigate this further we have generated several new antibodies to novel sites on p53 and mapped their epitopes using synthetic peptides. We find that antibodies to two other discrete sites in the core can also, like PAb240, recognize cryptic epitopes and distinguish mutant from wild-type conformations implying that the point mutations found in p53 in human tumours have widespread effects on the folding pattern of the DNA binding domain.
authors.affiliation: Department of Cellular and Molecular Oncology, Masaryk Memorial Cancer Institute, Brno, Czech Republic.
authors.forename: B
authors.initials: B
authors.lastname: Vojtesek
authors.forename: H
authors.initials: H
authors.lastname: Dolezalova
authors.forename: L
authors.initials: L
authors.lastname: Lauerova
authors.forename: M
authors.initials: M
authors.lastname: Svitakova
authors.forename: P
authors.initials: P
authors.lastname: Havlis
authors.forename: J
authors.initials: J
authors.lastname: Kovarik
authors.forename: C A
authors.initials: CA
authors.lastname: Midgley
authors.forename: D P
authors.initials: DP
authors.lastname: Lane
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D000939:Epitopes; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D011487:Protein Conformation; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D016159:Tumor Suppressor Protein p53