title: Radiolysis of lac repressor by gamma-rays and heavy ions: a two-hit model for protein inactivation.
abstract: Upon gamma-ray or argon ion irradiation of the lac repressor protein, its peptide chain is cleaved and the protein loses its lac operator-binding activity, as shown respectively by polyacrylamide gel electrophoresis and retardation gel electrophoresis. We developed phenomenological models that satisfactorily account for the experimental results: the peptide chain cleavage model considers that the average number of chain breaks per protomer is proportional to the irradiation dose and that the distribution of the number of breaks per protomer obeys Poisson's law. The repressor inactivation model takes into account the quaternary structure (a dimer of dimer) and the organization of the repressor in domains (two DNA binding sites, one per dimer). A protomer is inactivated by at least two different radiation-induced damages. A dimer is inactivated when at least one of the two protomers is inactivated. A tetramer is inactivated when both dimers are inactivated. From the combination of both models, we can deduce that chain cleavage cannot account for the protein inactivation, which should mainly result from oxidation of amino acid side chains. Indeed, particularly oxidizable and accessible amino acids (Tyr, His) are involved in the DNA binding process.
authors.affiliation: Centre de Biophysique Moléculaire, CNRS, 45071 Orléans Cedex 2, France. micharli@cnrs-orleans.fr
authors.forename: Michel
authors.initials: M
authors.lastname: Charlier
authors.forename: Séverine
authors.initials: S
authors.lastname: Eon
authors.forename: Edouard
authors.initials: E
authors.lastname: Sèche
authors.forename: Serge
authors.initials: S
authors.lastname: Bouffard
authors.forename: Françoise
authors.initials: F
authors.lastname: Culard
authors.forename: Mélanie
authors.initials: M
authors.lastname: Spotheim-Maurizot
mesh_terms: D001128:Argon; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000528:radiation effects*; D019281:Dimerization; D004926:Escherichia coli / Q000502:physiology / Q000528:radiation effects; D029968:Escherichia coli Proteins; D005720:Gamma Rays; D056968:Lac Repressors; D007785:Lactose / Q000037:antagonists & inhibitors*; D046911:Macromolecular Substances; D008954:Models, Biological; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry; D021122:Protein Subunits; D012097:Repressor Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000528:radiation effects*
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D056968:Lac Repressors; C539064:LacI protein, E coli; D046911:Macromolecular Substances; D010455:Peptides; D021122:Protein Subunits; D012097:Repressor Proteins; D001128:Argon; D007785:Lactose