title: Structural and functional characterization of 2-oxo-histidine in oxidized PerR protein.
abstract: In Bacillus subtilis, PerR is a metal-dependent sensor of hydrogen peroxide. PerR is a dimeric zinc protein with a regulatory site that coordinates either Fe(2+) (PerR-Zn-Fe) or Mn(2+) (PerR-Zn-Mn). Though most of the peroxide sensors use cysteines to detect H(2)O(2), it has been shown that reaction of PerR-Zn-Fe with H(2)O(2) leads to the oxidation of one histidine residue. Oxidation of PerR leads to the incorporation of one oxygen atom into His37 or His91. This study presents the crystal structure of the oxidized PerR protein (PerR-Zn-ox), which clearly shows a 2-oxo-histidine residue in position 37. Formation of 2-oxo-histidine is demonstrated and quantified by HPLC-MS/MS. EPR experiments indicate that PerR-Zn-H37ox retains a significant affinity for the regulatory metal, whereas PerR-Zn-H91ox shows a considerably reduced affinity for the metal ion. In spite of these major differences in terms of metal binding affinity, oxidation of His37 and/or His91 in PerR prevents DNA binding.
authors.affiliation: Commissariat à l'Energie Atomique, Institut de Recherches en Technologies et Sciences pour le Vivant, Laboratoire de Chimie et Biologie des Métaux, CEA-Grenoble, 17 avenue des Martyrs, 38054 Grenoble Cedex 9, France.
authors.forename: Daouda A K
authors.initials: DA
authors.lastname: Traoré
authors.forename: Abdelnasser
authors.initials: A
authors.lastname: El Ghazouani
authors.forename: Lilian
authors.initials: L
authors.lastname: Jacquamet
authors.forename: Franck
authors.initials: F
authors.lastname: Borel
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Ferrer
authors.forename: David
authors.initials: D
authors.lastname: Lascoux
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Ravanat
authors.forename: Michel
authors.initials: M
authors.lastname: Jaquinod
authors.forename: Geneviève
authors.initials: G
authors.lastname: Blondin
authors.forename: Christelle
authors.initials: C
authors.lastname: Caux-Thang
authors.forename: Victor
authors.initials: V
authors.lastname: Duarte
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Latour
mesh_terms: D001412:Bacillus subtilis / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D006639:Histidine / Q000737:chemistry* / Q000378:metabolism*; D013058:Mass Spectrometry; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011485:Protein Binding; D011487:Protein Conformation; D012097:Repressor Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012097:Repressor Proteins; C122138:peroxide repressor proteins; D006639:Histidine