Dataset Information

Functional implications of nitric oxide produced by mitochondria in mitochondrial metabolism.

ABSTRACT: The effects of endogenous production of NO., catalysed by the mitochondrial nitric oxide synthase (NOS), on mitochondrial metabolism were studied. The respiratory rates of intact mitochondria in State 4 were decreased by 40% and 28% with succinate and malate-glutamate, respectively, in the presence of L-arginine (L-Arg); conversely, the O2 uptake with NG-methyl-L-arginine (NMMA), a competitive inhibitor of NOS, was increased. The production of NO. and the inhibition of the respiratory rates were dependent on the metabolic state in which mitochondria were maintained: NO. production was probably supported by mitochondrial NADPH, the latter maintained by the energy-dependent transhydrogenase. In addition to the decline in the respiratory rate, an inhibition of ATP synthesis was also observed (40-50%) following supplementation with L-Arg. The dependence of the respiratory rates of mitochondria in State 3 and cytochrome oxidase activities on O2 concentrations with either L-Arg or NMMA indicated that both processes were competitively inhibited by NO. at the cytochrome oxidase level. This inhibition can be explained by the interaction of NO. with cytochrome oxidase at the binuclear centre. The role of NO. as a physiological modulator of cytochrome oxidase is discussed in terms of cellular metabolism.

SUBMITTER: Giulivi C

PROVIDER: S-EPMC1219527 | biostudies-other | 1998 Jun

REPOSITORIES: biostudies-other

ACCESS DATA

Similar Datasets

Project description:Melanoma patients experience inferior survival after biochemotherapy when their tumors contain numerous cells expressing the inducible isoform of NO synthase (iNOS) and elevated levels of nitrotyrosine, a product derived from NO. Although several lines of evidence suggest that NO promotes tumor growth and increases resistance to chemotherapy, it is unclear how it shapes these outcomes. Here we demonstrate that modulation of NO-mediated S-nitrosation of cellular proteins is strongly associated with the pattern of response to the anticancer agent cisplatin in human melanoma cells in vitro. Cells were shown to express iNOS constitutively, and to generate sustained nanomolar levels of NO intracellularly. Inhibition of NO synthesis or scavenging of NO enhanced cisplatin-induced apoptotic cell death. Additionally, pharmacologic agents disrupting S-nitrosation markedly increased cisplatin toxicity, whereas treatments favoring stabilization of S-nitrosothiols (SNOs) decreased its cytotoxic potency. Activity of the proapoptotic enzyme caspase-3 was higher in cells treated with a combination of cisplatin and chemicals that decreased NO/SNOs, whereas lower activity resulted from cisplatin combined with stabilization of SNOs. Constitutive protein S-nitrosation in cells was detected by analysis with biotin switch and reduction/chemiluminescence techniques. Moreover, intracellular NO concentration increased significantly in cells that survived cisplatin treatment, resulting in augmented S-nitrosation of caspase-3 and prolyl-hydroxylase-2, the enzyme responsible for targeting the prosurvival transcription factor hypoxia-inducible factor-1? for proteasomal degradation. Because activities of these enzymes are inhibited by S-nitrosation, our data thus indicate that modulation of intrinsic intracellular NO levels substantially affects cisplatin toxicity in melanoma cells. The underlying mechanisms may thus represent potential targets for adjuvant strategies to improve the efficacy of chemotherapy.

Dataset Information

Functional implications of nitric oxide produced by mitochondria in mitochondrial metabolism.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure