The Ubiquinone-binding Site in NADH:Ubiquinone Oxidoreductase from Escherichia coli

Gong, Xing; Xie, Tong; Yu, Linda; Hesterberg, Micaela; Scheide, Dierk; Friedrich, Thorsten; Yu, Chang An

doi:10.1074/jbc.m302361200

Cited by 71 publications

(57 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this case quinone binding sites and proton translocation machinery would be located close to the peripheral arm with its redox centers to allow direct interaction. This would be consistent with a location of subunits NuoL/M/N proximal to the peripheral arm and associated with the Q-binding pocket (29,32,33). In the second model, the catalytic module with electron carriers is an entity distinct from the proton pumping module and energy transduction takes place through long range conformational changes (4,25,36,37).…”

supporting

confidence: 66%

See 1 more Smart Citation

The Location of NuoL and NuoM Subunits in the Membrane Domain of the Escherichia coli Complex I

Holt

Morgan

Sazanov

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

The molecular organization of bacterial NADH: ubiquinone oxidoreductase (complex I or NDH-1) is not established, apart from a rough separation into dehydrogenase, connecting and membrane domains. In this work, complex I was purified from Escherichia coli and fragmented by replacing dodecylmaltoside with other detergents. Exchange into decyl maltoside led to the removal of the hydrophobic subunit NuoL from the otherwise intact complex. Diheptanoyl phosphocholine led to the loss of NuoL and NuoM subunits, whereas other subunits remained in the complex. The presence of N,Ndimethyldodecylamine N-oxide or Triton X-100 led to further disruption of the membrane domain into fragments containing NuoL/M/N, NuoA/K/N, and NuoH/J subunits. Among the hydrophilic subunits, NuoCD was most readily dissociated from the complex, whereas NuoB was partially dissociated from the peripheral arm assembly in N,N-dimethyldodecylamine N-oxide. A model of subunit arrangement in bacterial complex I based on these data is proposed. Subunits NuoL and NuoM, which are homologous to antiporters and are implicated in proton pumping, are located at the distal end of the membrane arm, spatially separated from the redox centers of the peripheral arm. This is consistent with proposals that the mechanism of proton pumping by complex I is likely to involve long range conformational changes.NADH:ubiquinone oxidoreductase (complex I, EC 1.6.5.3) is the first enzyme of the respiratory chains of most mitochondria and many bacteria. It catalyzes the transfer of two electrons from NADH to quinone, coupled to the translocation of about 4 protons across the membrane (for reviews, see Refs. 1-4). Complex I is one of the largest known membrane protein complexes. The bovine enzyme has a mass of ϳ980 kDa and is composed of about 46 subunits, including the seven hydrophobic ND subunits encoded in the mitochondrial genome (5). The simplest version of the complex in terms of protein content is the prokaryotic enzyme that has 13-14 subunits and a combined molecular mass of about 550 kDa (6). All of the subunits of bacterial complex I (also referred to as NDH-1) have analogues in the mitochondrial enzyme (4). Because of its relatively simple subunit composition, NDH-1 represents a useful "minimal" model to study the structure and function of complex I. It is currently the least understood component of the respiratory chain. In contrast to other enzymes of the respiratory chain, the atomic structure of complex I is not known and the mechanisms of proton pumping and electron transfer are not established.Electron microscopy has shown that both the mitochondrial and the bacterial enzyme have a characteristic L-shaped structure. One arm is embedded in the membrane and the other, the peripheral arm, protrudes into the mitochondrial matrix or bacterial cytoplasm. This was demonstrated at about 20 to 30 Å resolution for the enzyme from fungus Neurospora crassa (7, 8), yeast Yarrowia lipolytica (9), beef heart mitochondria (10), Escherichia coli (11,12), and the thermophile Aqu...

show abstract

supporting

confidence: 66%

“…NuoM by an azidoubiquinone derivative was not affected by NADH and inhibitors had a minor effect only, but this subunit was also proposed to be a part of a Q-binding pocket (33).…”

Section: Subunits Nuol and Nuom Are Removed From Complex I In Diheptamentioning

confidence: 96%

The Location of NuoL and NuoM Subunits in the Membrane Domain of the Escherichia coli Complex I

Holt

Morgan

Sazanov

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Cytochrome-c oxidase (CcO) activity was measured in the presence of phosphate buffer (50 mM, pH 7.4) and reduced cytochrome c (60 M; Sigma) (3). NADH-DH activity was measured in the presence of Tris-Cl buffer (20 mM, pH 8.0), NADH (150 M; Sigma), and coenzyme Q1 (100 M; Sigma) (8). Enzyme activity per milligram of protein in supernatant was calculated based on the millimolar extinction coefficient, ⑀ 550 nm ϭ 18.5 mM Ϫ1 ⅐cm Ϫ1 for cytochrome c and ⑀ 340 nm ϭ 6.22 mM Ϫ1 ⅐cm Ϫ1 for NADH.…”

Section: Animalsmentioning

confidence: 99%

Endothelial nitric oxide synthase (NOS3) knockout decreases NOS2 induction, limiting hyperoxygenation and conferring protection in the postischemic heart

Zhao¹,

Chen²,

He³

et al. 2007

American Journal of Physiology-Heart and Circulatory Physiology

View full text Add to dashboard Cite

Zhao X, Chen YR, He G, Zhang A, Druhan LJ, Strauch AR, Zweier JL. Endothelial nitric oxide synthase (NOS3) knockout decreases NOS2 induction, limiting hyperoxygenation and conferring protection in the postischemic heart. Am J Physiol Heart Circ Physiol 292: H1541-H1550, 2007. First published November 17, 2006; doi:10.1152/ajpheart.00264.2006.-Although it has been shown that endothelial nitric oxide synthase (eNOS)-derived nitric oxide downregulates mitochondrial oxygen consumption during early reperfusion, its effects on inducible NOS (iNOS) induction and myocardial injury during late reperfusion are unknown. Wild-type (WT) and eNOS Ϫ/Ϫ mice were subjected to 30 min of coronary ligation followed by reperfusion. Expression of iNOS mRNA and protein levels and peroxynitrite production were lower in postischemic myocardium of eNOS Ϫ/Ϫ mice than levels in WT mice 48 h postreperfusion. Significantly improved hemodynamics (ϮdP/dt, left ventricular systolic pressure, mean arterial pressure), increased rate pressure product, and reduced myocardial infarct size (18 Ϯ 2.5% vs. 31 Ϯ 4.6%) were found 48 h after reperfusion in eNOS Ϫ/Ϫ mice compared with WT mice. Myocardial infarct size was also significantly decreased in WT mice treated with the specific iNOS inhibitor 1400W (20.5 Ϯ 3.4%) compared with WT mice treated with PBS (33.9 Ϯ 5.3%). A marked reperfusion-induced hyperoxygenation state was observed by electron paramagnetic resonance oximetry in postischemic myocardium, but PO 2 values were significantly lower from 1 to 72 h in eNOS Ϫ/Ϫ than in WT mice. Cytochrome c-oxidase activity and NADH dehydrogenase activity were significantly decreased in postischemic myocardium in WT and eNOS Ϫ/Ϫ mice compared with baseline control, respectively, and NADH dehydrogenase activity was significantly higher in eNOS Ϫ/Ϫ than in WT mice. Thus deficiency of eNOS exerted a sustained beneficial effect on postischemic myocardium 48 h after reperfusion with preserved mitochondrial function, which appears to be due to decreased iNOS induction and decreased iNOS-derived peroxynitrite in postischemic myocardium.oxygen; superoxide; peroxynitrite; mitochondria; electron paramagnetic resonance; ischemia-reperfusion NITRIC OXIDE (NO) is an important modulator of cardiac performance and left ventricular (LV) remodeling after myocardial infarction. In the setting of ischemic preconditioning, there is evidence that endothelial nitric oxide synthase (eNOS)-derived NO is important for the acute window of protection as well as the subsequent induction of inducible NOS (iNOS), triggering the delayed window of protection (4). However, there has been considerable controversy regarding the effect of eNOS-and eNOS-derived NO on myocardial ischemia-reperfusion injury. In eNOS knockout mice (eNOS Ϫ/Ϫ

show abstract

“…22 NADH-DH activity was measured in the presence of Tris-HCl buffer (20 mmol/L, pH 8.0), NADH (150 mol/L, Sigma), and coenzyme Q 1 (100 mol/L, Sigma). 23 The extinction coefficients, ⑀ 550 nmϭ18.5 mmol/L Ϫ1 · cm Ϫ1 for cytochrome c and ⑀ 340 nmϭ6.22 mmol/L Ϫ1 · cm Ϫ1 for NADH, were used for activity calculation. Protein concentration of the tissue homogenate was measured by BCA assay (PIERCE Biotechnology).…”

Section: Activities Of Cco and Nadh-dhmentioning

confidence: 99%

Endothelium-Derived Nitric Oxide Regulates Postischemic Myocardial Oxygenation and Oxygen Consumption by Modulation of Mitochondrial Electron Transport

et al. 2005

View full text Add to dashboard Cite

Background-Nitric oxide (NO) production is increased in postischemic myocardium, and NO can control mitochondrial oxygen consumption in vitro. Therefore, we investigated the role of endothelial NO synthase (eNOS)-derived NO on in vivo regulation of oxygen consumption in the postischemic heart. Methods and Results-Mice were subjected to 30 minutes of coronary ligation followed by 60 minutes of reperfusion.Myocardial oxygen tension (PO 2 ) was monitored by electron paramagnetic resonance oximetry. In wild-type, N-nitro-L-arginine methyl ester (L-NAME)-treated (with 1 mg/mL in drinking water), and eNOS knockout (eNOS Ϫ/Ϫ ) mice, no difference was observed among baseline myocardial PO 2 values (8.6Ϯ0.7, 10.0Ϯ1.2, and 10.1Ϯ1.2 mm Hg, respectively) or those measured at 30 minutes of ischemia (1.4Ϯ0.6, 2.3Ϯ0.9, and 3.1Ϯ1.4 mm Hg, respectively). After reperfusion, myocardial PO 2 increased markedly (PϽ0.001 versus baseline in each group) but was much lower in L-NAME-treated and eNOS Ϫ/Ϫ mice (17.4Ϯ1.6 and 20.4Ϯ1.9 mm Hg) than in wild-type mice (46.5Ϯ1.7 mm Hg; PϽ0.001). A transient peak of myocardial PO 2 was observed at early reperfusion in wild-type mice. No reactive hyperemia was observed during early reperfusion. Endothelial NO decreased the rate-pressure product (PϽ0.05), upregulated cytochrome c oxidase (CcO) mRNA expression (PϽ0.01) with no change in CcO activity, and inhibited NADH dehydrogenase (NADH-DH) activity (PϽ0.01) without alteration of NADH-DH mRNA expression. Peroxynitrite-mediated tyrosine nitration was higher in hearts from wild-type mice than in eNOS Ϫ/Ϫ or L-NAME-treated hearts. Conclusions-eNOS-derived

show abstract

The Ubiquinone-binding Site in NADH:Ubiquinone Oxidoreductase from Escherichia coli

Cited by 71 publications

References 42 publications

The Location of NuoL and NuoM Subunits in the Membrane Domain of the Escherichia coli Complex I

The Location of NuoL and NuoM Subunits in the Membrane Domain of the Escherichia coli Complex I

Endothelial nitric oxide synthase (NOS3) knockout decreases NOS2 induction, limiting hyperoxygenation and conferring protection in the postischemic heart

Endothelium-Derived Nitric Oxide Regulates Postischemic Myocardial Oxygenation and Oxygen Consumption by Modulation of Mitochondrial Electron Transport

Contact Info

Product

Resources

About