The cellular production of hydrogen peroxide

作者：Alberto Boveris, Nozomu Oshino, Britton Chance · 发表于：Biochemical Journal · 年份：1972 · DOI：10.1042/bj1280617 · 被引用次数：1551 · 研究领域：Metabolomics and Mass Spectrometry Studies、Mitochondrial Function and Pathology、Adipose Tissue and Metabolism

1. The enzyme-substrate complex of yeast cytochrome c peroxidase is used as a sensitive, specific and accurate spectrophotometric H(2)O(2) indicator. 2. The cytochrome c peroxidase assay is suitable for use with subcellular fractions from tissue homogenates as well as with pure enzyme systems to measure H(2)O(2) generation. 3. Mitochondrial substrates entering the respiratory chain on the substrate side of the antimycin A-sensitive site support the mitochondrial generation of H(2)O(2). Succinate, the most effective substrate, yields H(2)O(2) at a rate of 0.5nmol/min per mg of protein in state 4. H(2)O(2) generation is decreased in the state 4-->state 3 transition. 4. In the combined mitochondrial-peroxisomal fraction of rat liver the changes in the mitochondrial generation of H(2)O(2) modulated by substrate, ADP and antimycin A are followed by parallel changes in the saturation of the intraperoxisomal catalase intermediate. 5. Peroxisomes supplemented with uric acid generate extraperoxisomal H(2)O(2) at a rate (8.6-16.4nmol/min per mg of protein) that corresponds to 42-61% of the rate of uric acid oxidation. Addition of azide increases these H(2)O(2) rates by a factor of 1.4-1.7. 6. The concentration of cytosolic uric acid is shown to vary during the isolation of the cellular fractions. 7. Microsomal fractions produce H(2)O(2) (up to 1.7nmol/min per mg of protein) at a ratio of 0.71-0.86mol of H(2)O(2)/mol of NADP(+) during the oxidation of NADPH. H(2)O(2) is also generated (...