title: Scavenging of H2O2 by mouse brain mitochondria.
abstract: Mitochondrial reactive oxygen species (ROS) metabolism is unique in that mitochondria both generate and scavenge ROS. Recent estimates of ROS scavenging capacity of brain mitochondria are surprisingly high, ca. 9-12 nmol H2O2/min/mg, which is ~100 times higher than the rate of ROS generation. This raises a question whether brain mitochondria are a source or a sink of ROS. We studied the interaction between ROS generation and scavenging in mouse brain mitochondria by measuring the rate of removal of H2O2 added at a concentration of 0.4 μM, which is close to the reported physiological H2O2 concentrations in tissues, under conditions of low and high levels of mitochondrial H2O2 generation. With NAD-linked substrates, the rate of H2O2 generation by mitochondria was ~50-70 pmol/min/mg. The H2O2 scavenging dynamics was best approximated by the first order reaction equation. H2O2 scavenging was not affected by the uncoupling of mitochondria, phosphorylation of added ADP, or the genetic ablation of glutathione peroxidase 1, but decreased in the absence of respiratory substrates, in the presence of thioredoxin reductase inhibitor auranofin, or in partially disrupted mitochondria. With succinate, the rate of H2O2 generation was ~2,200-2,900 pmol/min/mg; the scavenging of added H2O2 was masked by a significant accumulation of generated H2O2 in the assay medium. The obtained data were fitted into a simple model that reasonably well described the interaction between H2O2 scavenging and production. It showed that mitochondria are neither a sink nor a source of H2O2, but can function as both at the same time, efficiently stabilizing exogenous H2O2 concentration at a level directly proportional to the ratio of the H2O2 generation rate to the rate constant of the first order scavenging reaction.
authors.affiliation: Brain and Mind Research Institute, Weill Cornell Medical College, 407 East 61st Street, 5th floor, New York, NY, 10065, USA, ans2024@med.cornell.edu.
authors.forename: Anatoly A
authors.initials: AA
authors.lastname: Starkov
authors.forename: Alexander Yu
authors.initials: AY
authors.lastname: Andreyev
authors.forename: Steven F
authors.initials: SF
authors.lastname: Zhang
authors.forename: Natalia N
authors.initials: NN
authors.lastname: Starkova
authors.forename: Maria
authors.initials: M
authors.lastname: Korneeva
authors.forename: Mikhail
authors.initials: M
authors.lastname: Syromyatnikov
authors.forename: Vasily N
authors.initials: VN
authors.lastname: Popov
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D018384:Oxidative Stress; D017382:Reactive Oxygen Species
chemical_list: D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide