title: The oxidation produced by hydrogen peroxide on Ca-ATP-G-actin.
abstract: We report here that in vitro exposure of monomeric actin to hydrogen peroxide leads to a conversion of 6 of the 16 methionine residues to methionine sulfoxide residues. Although the initial effect of H2O2 on actin is the oxidation of Cys374, we have found that Met44, Met47, Met176, Met190, Met269, and Met355 are the other sites of the oxidative modification. Met44 and Met47 are the methionyl sites first oxidized. The methionine residues that are oxidized are not simply related to their accessibility to the external medium and are found in all four subdomains of actin. The conformations of subdomain 1, a region critical for the functional binding of different actin-binding proteins, and subdomain 2, which plays important roles in the polymerization process and stabilization of the actin filament, are changed upon oxidation. The conformational changes are deduced from the increased exposure of hydrophobic residues, which correlates with methionine sulfoxide formation, from the perturbations in tryptophan fluorescence, and from the decreased susceptibility to limited proteolysis of oxidized actin.
authors.affiliation: Department of Biology, University of Milan, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Milzani
authors.forename: R
authors.initials: R
authors.lastname: Rossi
authors.forename: P
authors.initials: P
authors.lastname: Di Simplicio
authors.forename: D
authors.initials: D
authors.lastname: Giustarini
authors.forename: R
authors.initials: R
authors.lastname: Colombo
authors.forename: I
authors.initials: I
authors.lastname: DalleDonne
mesh_terms: D000199:Actins / Q000737:chemistry*; D000255:Adenosine Triphosphate / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D006861:Hydrogen Peroxide / Q000737:chemistry*; D008715:Methionine / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011817:Rabbits
chemical_list: D000199:Actins; D000255:Adenosine Triphosphate; D008715:Methionine; D006861:Hydrogen Peroxide