Poderoso J J, Lisdero C, Schöpfer F, Riobó N, Carreras M C, Cadenas E, Boveris A
Laboratory of Oxygen Metabolism, University Hospital, School of Medicine, University of Buenos Aires, Córdoba 2351, 1120 Buenos Aires,
J Biol Chem. 1999 Dec 31;274(53):37709-16. doi: 10.1074/jbc.274.53.37709.
The reversible inhibitory effects of nitric oxide (.NO) on mitochondrial cytochrome oxidase and O(2) uptake are dependent on intramitochondrial.NO utilization. This study was aimed at establishing the mitochondrial pathways for.NO utilization that regulate O-(2) generation via reductive and oxidative reactions involving ubiquinol oxidation and peroxynitrite (ONOO(-)) formation. For this purpose, experimental models consisting of intact mitochondria, ubiquinone-depleted/reconstituted submitochondrial particles, and ONOO(-)-supplemented mitochondrial membranes were used. The results obtained from these experimental approaches strongly suggest the occurrence of independent pathways for.NO utilization in mitochondria, which effectively compete with the binding of.NO to cytochrome oxidase, thereby releasing this inhibition and restoring O(2) uptake. The pathways for.NO utilization are discussed in terms of the steady-state levels of.NO and O-(2) and estimated as a function of O(2) tension. These calculations indicate that mitochondrial.NO decays primarily by pathways involving ONOO(-) formation and ubiquinol oxidation and, secondarily, by reversible binding to cytochrome oxidase.
一氧化氮(·NO)对线粒体细胞色素氧化酶和氧气摄取的可逆抑制作用取决于线粒体内·NO的利用情况。本研究旨在确定·NO利用的线粒体途径,该途径通过涉及泛醇氧化和过氧亚硝酸盐(ONOO⁻)形成的还原和氧化反应来调节超氧阴离子(O₂⁻)的产生。为此,使用了由完整线粒体、泛醌耗尽/重组的亚线粒体颗粒和补充了ONOO⁻的线粒体膜组成的实验模型。从这些实验方法获得的结果强烈表明,线粒体中存在独立的·NO利用途径,这些途径有效地与·NO与细胞色素氧化酶的结合竞争,从而解除这种抑制并恢复氧气摄取。根据·NO和O₂⁻的稳态水平讨论了·NO利用途径,并将其估计为氧气张力的函数。这些计算表明,线粒体·NO主要通过涉及ONOO⁻形成和泛醇氧化的途径衰减,其次通过与细胞色素氧化酶的可逆结合衰减。