title: Oxidation of Са2+-Binding Domain of NADPH Oxidase 5 (NOX5): Toward Understanding the Mechanism of Inactivation of NOX5 by ROS.
abstract: NOX5 protein, one of the most active generators of reactive oxygen species (ROS), plays an important role in many processes, including regulation of cell growth, death and differentiation. Because of its central role in ROS generation, it needs to be tightly regulated to guarantee cellular homeostasis. Contrary to other members of NADPH-oxidases family, NOX5 has its own regulatory calcium-binding domain and thus could be activated directly by calcium ions. While several mechanisms of activation have been described, very little is known about the mechanisms that could prevent the overproduction of ROS by NOX5. In the present study using calorimetric methods and circular dichroism we found that oxidation of cysteine and methionine residues of NOX5 decreases binding of Ca2+ ions and perturbs both secondary and tertiary structure of protein. Our data strongly suggest that oxidation of calcium-binding domain of NOX5 could be implicated in its inactivation, serving as a possible defense mechanism against oxidative stress.
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Street 32, 119991 Moscow, Russia.
authors.forename: Irina Yu
authors.initials: IY
authors.lastname: Petrushanko
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Street 32, 119991 Moscow, Russia.
authors.forename: Vladimir M
authors.initials: VM
authors.lastname: Lobachev
authors.affiliation: Moscow Institute of Physics and Technology, 141700 Dolgoprudnyi, Moscow Region, Russia.
authors.forename: Alexey S
authors.initials: AS
authors.lastname: Kononikhin
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Street 32, 119991 Moscow, Russia.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Makarov
authors.affiliation: Aix-Marseille University, Inserm, CRO2 UMR_S 911, Faculté de Pharmacie, 13385 Marseille, France.
authors.forename: Francois
authors.initials: F
authors.lastname: Devred
authors.affiliation: Aix-Marseille University, Inserm, CRO2 UMR_S 911, Faculté de Pharmacie, 13385 Marseille, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Kovacic
authors.affiliation: Institute of General Pathology and Pathophysiology, RAMS, 125315, Moscow, Russian Federation.
authors.forename: Aslan A
authors.initials: AA
authors.lastname: Kubatiev
authors.affiliation: Aix-Marseille University, Inserm, CRO2 UMR_S 911, Faculté de Pharmacie, 13385 Marseille, France.|Institute of General Pathology and Pathophysiology, RAMS, 125315, Moscow, Russian Federation.
authors.forename: Philipp O
authors.initials: PO
authors.lastname: Tsvetkov
mesh_terms: D000595:Amino Acid Sequence; D002151:Calorimetry; D002152:Calorimetry, Differential Scanning; D002853:Chromatography, Liquid; D002942:Circular Dichroism; D003545:Cysteine / Q000378:metabolism; D006706:Homeostasis; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D000074664:NADPH Oxidase 5; D019255:NADPH Oxidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D000072417:Protein Domains; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D053719:Tandem Mass Spectrometry
chemical_list: D008565:Membrane Proteins; D017382:Reactive Oxygen Species; D008715:Methionine; D000074664:NADPH Oxidase 5; D019255:NADPH Oxidases; C431571:NOX5 protein, human; D003545:Cysteine