Nitric oxide inhibits mitochondrial NADH:ubiquinone reductase activity through peroxynitrite formation

2001 ◽  
Vol 359 (1) ◽  
pp. 139-145 ◽  
Author(s):  
Natalia A. RIOBÓ ◽  
Emilio CLEMENTI ◽  
Mariana MELANI ◽  
Alberto BOVERIS ◽  
Enrique CADENAS ◽  
...  
Keyword(s):  
Complex I ◽  
Superoxide Radical ◽  
Reductase Activity ◽  
Complex Iii ◽  
Submitochondrial Particles ◽  
Nadh:Ubiquinone Reductase ◽  
Complex I Activity ◽  
State Concentration ◽  
The Brain

This study was aimed at assessing the effects of long-term exposure to NO of respiratory activities in mitochondria from different tissues (with different ubiquinol contents), under conditions that either promote or prevent the formation of peroxynitrite. Mitochondria and submitochondrial particles isolated from rat heart, liver and brain were exposed either to a steady-state concentration or to a bolus addition of NO. NO induced the mitochondrial production of superoxide anions, hydrogen peroxide and peroxynitrite, the latter shown by nitration of mitochondrial proteins. Long-term incubation of mitochondrial membranes with NO resulted in a persistent inhibition of NADH:cytochrome c reductase activity, interpreted as inhibition of NADH:ubiquinone reductase (Complex I) activity, whereas succinate:cytochrome c reductase activity, including Complex II and Complex III electron transfer, remained unaffected. This selective effect of NO and derived species was partially prevented by superoxide dismutase and uric acid. In addition, peroxynitrite mimicked the effect of NO, including tyrosine nitration of some Complex I proteins. These results seem to indicate that the inhibition of NADH:ubiquinone reductase (Complex I) activity depends on the NO-induced generation of superoxide radical and peroxynitrite and that Complex I is selectively sensitive to peroxynitrite. Inhibition of Complex I activity by peroxynitrite may have critical implications for energy supply in tissues such as the brain, whose mitochondrial function depends largely on the channelling of reducing equivalents through Complex I.

scholarly journals Isoflurane Differentially Modulates Mitochondrial Reactive Oxygen Species Production via  Forward versus  Reverse Electron Transport Flow

2011 ◽  
Vol 115 (3) ◽  
pp. 531-540 ◽  
Author(s):  
Naoyuki Hirata ◽  
Yon Hee Shim ◽  
Danijel Pravdic ◽  
Nicole L. Lohr ◽  
Philip F. Pratt ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Electron Transport ◽  
Complex I ◽  
Ros Generation ◽  
Ros Production ◽  
Oxygen Species ◽  
Submitochondrial Particles ◽  
Anesthetic Preconditioning ◽  
Cardioprotective Effects ◽  
Complex I Activity

Background Reactive oxygen species (ROS) mediate the effects of anesthetic precondition to protect against ischemia and reperfusion injury, but the mechanisms of ROS generation remain unclear. In this study, the authors investigated if mitochondria-targeted antioxidant (mitotempol) abolishes the cardioprotective effects of anesthetic preconditioning. Further, the authors investigated the mechanism by which isoflurane alters ROS generation in isolated mitochondria and submitochondrial particles. Methods Rats were pretreated with 0.9% saline, 3.0 mg/kg mitotempol in the absence or presence of 30 min exposure to isoflurane. Myocardial infarction was induced by left anterior descending artery occlusion for 30 min followed by reperfusion for 2 h and infarct size measurements. Mitochondrial ROS production was determined spectrofluorometrically. The effect of isoflurane on enzymatic activity of mitochondrial respiratory complexes was also determined. Results Isoflurane reduced myocardial infarct size (40 ± 9% = mean ± SD) compared with control experiments (60 ± 4%). Mitotempol abolished the cardioprotective effects of anesthetic preconditioning (60 ± 9%). Isoflurane enhanced ROS generation in submitochondrial particles with nicotinamide adenine dinucleotide (reduced form), but not with succinate, as substrate. In intact mitochondria, isoflurane enhanced ROS production in the presence of rotenone, antimycin A, or ubiquinone when pyruvate and malate were substrates, but isoflurane attenuated ROS production when succinate was substrate. Mitochondrial respiratory experiments and electron transport chain complex assays revealed that isoflurane inhibited only complex I activity. Conclusions The results demonstrated that isoflurane produces ROS at complex I and III of the respiratory chain via the attenuation of complex I activity. The action on complex I decreases unfavorable reverse electron flow and ROS release in myocardium during reperfusion.

scholarly journals Global ischaemia induces a biphasic response of the mitochondrial respiratory chain. Anoxic pre-perfusion protects against ischaemic damage

1992 ◽  
Vol 281 (3) ◽  
pp. 709-715 ◽  
Author(s):  
K Veitch ◽  
A Hombroeckx ◽  
D Caucheteux ◽  
H Pouleur ◽  
L Hue
Keyword(s):  
Respiratory Chain ◽  
Complex I ◽  
Nadh Oxidase ◽  
Mitochondrial Respiratory Chain ◽  
Complex Iii ◽  
Biphasic Response ◽  
Ischaemic Damage ◽  
Global Ischaemia ◽  
Complex I Activity ◽  
Mitochondrial Respiratory

Studies of Langendorff-perfused rat hearts have revealed a biphasic response of the mitochondrial respiratory chain to global ischaemia. The initial effect is a 30-40% increase in the rate of glutamate/malate oxidation after 10 min of ischaemia, owing to an increase in the capacity for NADH oxidation. This effect is followed by a progressive decrease in these oxidative activities as the ischaemia is prolonged, apparently owing to damage to Complex I at a site subsequent to the NADH dehydrogenase component. This damage is exacerbated by reperfusion, which causes a further decrease in Complex I activity and also decreases the activities of the other complexes, most notably of Complex III. Perfusion for up to 1 h with anoxic buffer produced only the increase in NADH oxidase activity, and neither anoxia alone, nor anoxia and reperfusion, caused loss of Complex I activity. Perfusing for 3-10 min with anoxic buffer before 1 h of global ischaemia had a significant protective effect against the ischaemia-induced damage to Complex I.

scholarly journals Coenzyme Q1 redox metabolism during passage through the rat pulmonary circulation and the effect of hyperoxia

2008 ◽  
Vol 105 (4) ◽  
pp. 1114-1126 ◽  
Author(s):  
Said H. Audi ◽  
Marilyn P. Merker ◽  
Gary S. Krenz ◽  
Taniya Ahuja ◽  
David L. Roerig ◽  
...  
Keyword(s):  
Pulmonary Circulation ◽  
Complex I ◽  
Dominant Role ◽  
Coenzyme Q ◽  
Redox Status ◽  
Complex Iii ◽  
Mitochondrial Complex ◽  
Quinone Oxidoreductase ◽  
Arterial Inflow ◽  
Complex I Activity

The objective was to evaluate the pulmonary disposition of the ubiquinone homolog coenzyme Q1 (CoQ1) on passage through lungs of normoxic (exposed to room air) and hyperoxic (exposed to 85% O2 for 48 h) rats. CoQ1 or its hydroquinone (CoQ1H2) was infused into the arterial inflow of isolated, perfused lungs, and the venous efflux rates of CoQ1H2 and CoQ1 were measured. CoQ1H2 appeared in the venous effluent when CoQ1 was infused, and CoQ1 appeared when CoQ1H2 was infused. In normoxic lungs, CoQ1H2 efflux rates when CoQ1 was infused decreased by 58 and 33% in the presence of rotenone (mitochondrial complex I inhibitor) and dicumarol [NAD(P)H-quinone oxidoreductase 1 (NQO1) inhibitor], respectively. Inhibitor studies also revealed that lung CoQ1H2 oxidation was via mitochondrial complex III. In hyperoxic lungs, CoQ1H2 efflux rates when CoQ1 was infused decreased by 23% compared with normoxic lungs. Based on inhibitor effects and a kinetic model, the effect of hyperoxia could be attributed predominantly to 47% decrease in the capacity of complex I-mediated CoQ1 reduction, with no change in the other redox processes. Complex I activity in lung homogenates was also lower for hyperoxic than for normoxic lungs. These studies reveal that lung complexes I and III and NQO1 play a dominant role in determining the vascular concentration and redox status of CoQ1 during passage through the pulmonary circulation, and that exposure to hyperoxia decreases the overall capacity of the lung to reduce CoQ1 to CoQ1H2 due to a depression in complex I activity.

scholarly journals Natural substances (acetogenins) from the family Annonaceae are powerful inhibitors of mitochondrial NADH dehydrogenase (Complex I)

1994 ◽  
Vol 301 (1) ◽  
pp. 161-167 ◽  
Author(s):  
M Degli Esposti ◽  
A Ghelli ◽  
M Ratta ◽  
D Cortes ◽  
E Estornell
Keyword(s):  
Complex I ◽  
Nadh Dehydrogenase ◽  
Potent Inhibitor ◽  
Submitochondrial Particles ◽  
Annonaceous Acetogenins ◽  
Natural Substances ◽  
Mammalian Mitochondria ◽  
The Family ◽  
Complex I Activity ◽  
First Time

Natural products from the plants of the family Annonaceae, collectively called Annonaceous acetogenins, are very potent inhibitors of the NADH-ubiquinone reductase (Complex I) activity of mammalian mitochondria. The properties of five of such acetogenins are compared with those of rotenone and piericidin, classical potent inhibitors of Complex I. Rolliniastatin-1 and rolliniastatin-2 are more powerful than piericidin in terms of both their inhibitory constant and the protein-dependence of their titre in bovine submitochondrial particles. These acetogenins could be considered therefore the most potent inhibitors of mammalian Complex I. Squamocin and otivarin also have an inhibitory constant lower than that of piericidin, but display a larger protein-dependence of the titre. Squamocin and otivarin, contrary to the other acetogenins, behave qualitatively like rotenone. Rolliniastatin-2 shows unique properties as its interaction, although mutually exclusive to that of piericidin, appears to be mutually non-exclusive to that of rotenone. It is the first time that a potent inhibitor of Complex I is found not to overlap the active site of rotenone.

scholarly journals Adverse effects of anti-tuberculosis drugs on HepG2 cell bioenergetics

2016 ◽  
Vol 36 (6) ◽  
pp. 616-625 ◽  
Author(s):  
E Elmorsy ◽  
SM Attalla ◽  
E Fikry ◽  
A Kocon ◽  
R Turner ◽  
...  
Keyword(s):  
Complex I ◽  
Drug Exposure ◽  
Lactate Production ◽  
Complex Iii ◽  
Mitochondrial Complex I ◽  
Mitochondrial Complex ◽  
Drug Induced ◽  
Atp Levels ◽  
Drug Treatments ◽  
Complex I Activity

Tuberculosis (TB) is an intractable chronic infection. Disease treatment with anti-TB drugs remains challenging due to drug-induced hepatotoxicity. The toxicity of the anti-TB drugs rifampicin (RIF), isoniazid (INH) and pyrazinamide (PZA) either alone or in combination was investigated in HepG2 cells. Assays of intracellular adenosine triphosphate (ATP) levels at 4-, 24- and 48-h post-exposure to gradient concentrations of RIF, INH and PZA were conducted. Drug-induced effects on mitochondrial membrane potential (MMP), mitochondrial complex I and complex III activity, nicotinamide adenine dinucleotide (NAD+) levels and cellular lactate production were assessed. Decreased ATP levels were dose-dependent and correlated with drug exposure duration. Approximate 24-h IC50s were 0.5 mM, 70 mM and 84 mM for RIF, INH and PZA, respectively. Twenty-four hours post-drug treatment, reductions of MMP ( p = 0.0005), mitochondrial complex I and III activities ( p = 0.0001 and p = 0.0003, respectively), NAD+ levels ( p = 0.0057) and increased lactate production ( p < 0.0001) were observed. Drug combinations used to mimic cumulative drug treatments induced a synergistic inhibition of mitochondrial complex I activity. An assessment of cellular ultrastructure using transmission electron microscopy indicated drug-induced mitophagy. Collectively, our study suggests that hepatotoxicity of commonly employed anti-TB drugs is mediated by their curtailment of mitochondrial function.

Preservation of complex I function during hypoxia-reoxygenation-induced mitochondrial injury in proximal tubules

2004 ◽  
Vol 286 (4) ◽  
pp. F749-F759 ◽  
Author(s):  
Thorsten Feldkamp ◽  
Andreas Kribben ◽  
Nancy F. Roeser ◽  
Ruth A. Senter ◽  
Sarah Kemner ◽  
...  
Keyword(s):  
Complex I ◽  
Reductase Activity ◽  
Ischemia Reperfusion ◽  
Proximal Tubules ◽  
Progressive Reduction ◽  
Extensive Degradation ◽  
Complex I Activity ◽  
Insight Into ◽  
Hypoxia Reoxygenation

Inhibition of complex I has been considered to be an important contributor to mitochondrial dysfunction in tissues subjected to ischemia-reperfusion. We have investigated the role of complex I in a severe energetic deficit that develops in kidney proximal tubules subjected to hypoxia-reoxygenation and is strongly ameliorated by supplementation with specific citric acid cycle metabolites, including succinate and the combination of α-ketoglutarate plus malate. NADH: ubiquinone reductase activity in the tubules was decreased by only 26% during 60-min hypoxia and did not change further during 60-min reoxygenation. During titration of complex I activity with rotenone, progressive reduction of NAD+ to NADH was detected at >20% complex I inhibition, but substantial decreases in ATP levels and mitochondrial membrane potential did not occur until >70% inhibition. NAD+ was reduced to NADH during hypoxia, but the NADH formed was fully reoxidized during reoxygenation, consistent with the conclusion that complex I function was not limiting for recovery. Extensive degradation of cytosolic and mitochondrial NAD(H) pools occurred during either hypoxia or severe electron transport inhibition by rotenone, with patterns of metabolite accumulation consistent with catabolism by both NAD+ glycohydrolase and pyrophosphatase. This degradation was strongly blocked by α-ketoglutarate plus malate. The data demonstrate surprisingly little sensitivity of these cells to inhibition of complex I and high levels of resistance to development of complex I dysfunction during hypoxia-reoxygenation and indicate that events upstream of complex I are important for the energetic deficit. The work provides new insight into fundamental aspects of mitochondrial pathophysiology in proximal tubules during acute renal failure.

scholarly journals Lipid peroxidation and the reduction of ADP-Fe3+ chelate by NADH-ubiquinone reductase preparation from bovine heart mitochondria

1980 ◽  
Vol 192 (3) ◽  
pp. 861-866 ◽  
Author(s):  
K Takeshige ◽  
R Takayanagi ◽  
S Minakami
Keyword(s):  
Lipid Peroxidation ◽  
Complex I ◽  
Nadh Dehydrogenase ◽  
Reductase Activity ◽  
Thiol Group ◽  
Submitochondrial Particles ◽  
Essential Step ◽  
Sensitive Site ◽  
Mitochondrial Lipids ◽  
Optimal Ph

The NADH-ubiquinone reductase preparation (Complex I) of bovine hart mitochondria catalysed in the presence of reduced coenzymes and ADP-Fe3+ the lipid peroxidation of liposomes prepared from mitochondrial lipids. The apparent Km values for the coenzymes and the optimal pH of the reactions agreed well with those of the lipid peroxidation of the submitochondrial particles treated with rotenone. On assay of the reduction of ADP-Fe3+ chelate by the reduction of cytochrome c in the presence of superoxide dismutase and antimycin A or by the oxidation of reduced coenzymes, the reactions were not affected by rotenone but were inhibited by thiol-group inhibitors. The properties of the ADP-Fe3+ reductase activity were highly consistent with those of the lipid-peroxidation reaction. These observations suggest that electrons from reduced coenzymes are transferred to ADP-Fe3+ chelate from a component between a mercurial-sensitive site and the rotenone-sensitive one of the NADH dehydrogenase and that the reduction of ADP-Fe3+ chelate by the NADH dehydrogenase is an essential step in the lipid peroxidation.

scholarly journals Differential responses of targeted lung redox enzymes to rat exposure to 60 or 85% oxygen

2011 ◽  
Vol 111 (1) ◽  
pp. 95-107 ◽  
Author(s):  
Zhuohui Gan ◽  
David L. Roerig ◽  
Anne V. Clough ◽  
Said H. Audi
Keyword(s):  
Lung Tissue ◽  
Complex I ◽  
Complex Iii ◽  
Oxygen Species ◽  
Mitochondrial Complex ◽  
Complex Iv ◽  
Arterial Inflow ◽  
Tissue Homogenates ◽  
Species Production ◽  
Complex I Activity

Rat exposure to 60% O2 (hyper-60) or 85% O2 (hyper-85) for 7 days confers susceptibility or tolerance, respectively, of the otherwise lethal effects of exposure to 100% O2. The objective of this study was to determine whether activities of the antioxidant cytosolic enzyme NAD(P)H:quinone oxidoreductase 1 (NQO1) and mitochondrial complex III are differentially altered in hyper-60 and hyper-85 lungs. Duroquinone (DQ), an NQO1 substrate, or its hydroquinone (DQH2), a complex III substrate, was infused into the arterial inflow of isolated, perfused lungs, and the venous efflux rates of DQH2 and DQ were measured. Based on inhibitor effects and kinetic modeling, capacities of NQO1-mediated DQ reduction ( Vmax1) and complex III-mediated DQH2 oxidation ( Vmax2) increased by ∼140 and ∼180% in hyper-85 lungs, respectively, compared with rates in lungs of rats exposed to room air (normoxic). In hyper-60 lungs, Vmax1 increased by ∼80%, with no effect on Vmax2. Additional studies revealed that mitochondrial complex I activity in hyper-60 and hyper-85 lung tissue homogenates was ∼50% lower than in normoxic lung homogenates, whereas mitochondrial complex IV activity was ∼90% higher in only hyper-85 lung tissue homogenates. Thus NQO1 activity increased in both hyper-60 and hyper-85 lungs, whereas complex III activity increased in hyper-85 lungs only. This increase, along with the increase in complex IV activity, may counter the effects the depression in complex I activity might have on tissue mitochondrial function and/or reactive oxygen species production and may be important to the tolerance of 100% O2 observed in hyper-85 rats.

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