title: Nitric oxide inhibits succinate dehydrogenase-driven oxygen consumption in potato tuber mitochondria in an oxygen tension-independent manner.
abstract: NO (nitric oxide) is described as an inhibitor of plant and mammalian respiratory chains owing to its high affinity for COX (cytochrome c oxidase), which hinders the reduction of oxygen to water. In the present study we show that in plant mitochondria NO may interfere with other respiratory complexes as well. We analysed oxygen consumption supported by complex I and/or complex II and/or external NADH dehydrogenase in Percoll-isolated potato tuber (Solanum tuberosum) mitochondria. When mitochondrial respiration was stimulated by succinate, adding the NO donors SNAP (S-nitroso-N-acetyl-DL-penicillamine) or DETA-NONOate caused a 70% reduction in oxygen consumption rate in state 3 (stimulated with 1 mM of ADP). This inhibition was followed by a significant increase in the Km value of SDH (succinate dehydrogenase) for succinate (Km of 0.77±0.19 to 34.3±5.9 mM, in the presence of NO). When mitochondrial respiration was stimulated by external NADH dehydrogenase or complex I, NO had no effect on respiration. NO itself and DETA-NONOate had similar effects to SNAP. No significant inhibition of respiration was observed in the absence of ADP. More importantly, SNAP inhibited PTM (potato tuber mitochondria) respiration independently of oxygen tensions, indicating a different kinetic mechanism from that observed in mammalian mitochondria. We also observed, in an FAD reduction assay, that SNAP blocked the intrinsic SDH electron flow in much the same way as TTFA (thenoyltrifluoroacetone), a non-competitive SDH inhibitor. We suggest that NO inhibits SDH in its ubiquinone site or its Fe-S centres. These data indicate that SDH has an alternative site of NO action in plant mitochondria.
authors.affiliation: Laboratório de Bioenergética e Fisiologia Mitocondrial, Programa de Biofísica e Bioquímica Celular, Instituto de Bioquímica Médica, Centro de Ciências da Saúde, Universidade Federal do Rio de Janeiro, Rio de Janeiro, CEP 21941-590, Brazil.
authors.forename: Vagner
authors.initials: V
authors.lastname: Simonin
authors.forename: Antonio
authors.initials: A
authors.lastname: Galina
mesh_terms: D000818:Animals; D001923:Brain Chemistry / Q000502:physiology; D051379:Mice; D008928:Mitochondria / Q000201:enzymology / Q000502:physiology*; D008930:Mitochondria, Liver / Q000201:enzymology / Q000502:physiology; D009569:Nitric Oxide / Q000737:chemistry / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000737:chemistry / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption / Q000502:physiology*; D011198:Solanum tuberosum / Q000201:enzymology / Q000502:physiology*; D013367:Submitochondrial Particles / Q000201:enzymology / Q000502:physiology*; D013385:Succinate Dehydrogenase / Q000037:antagonists & inhibitors* / Q000502:physiology
chemical_list: D020030:Nitric Oxide Donors; D009569:Nitric Oxide; D013385:Succinate Dehydrogenase; D010100:Oxygen