Abstract
Bovine NADH:ubiquinone oxidoreductase (Complex I) is the first complex in the mitochondrial respiratory chain. It has long been assumed that it contained only one FMN group. However, as demonstrated in 2003, the intact enzyme contains two FMN groups. The second FMN was proposed to be located in a conserved flavodoxin fold predicted to be present in the PSST subunit. The long-known reaction of Complex I with NADPH differs in many aspects from that with NADH. It was proposed that the second flavin group was specifically involved in the reaction with NADPH. The X-ray structure of the hydrophilic domain of Complex I from Thermus thermophilus (Sazanov and Hinchliffe 2006, Science 311, 1430–1436) disclosed the positions of all redox groups of that enzyme and of the subunits holding them. The PSST subunit indeed contains the predicted flavodoxin fold although it did not contain FMN. Inspired by this structure, the present paper describes a re-evaluation of the enigmatic reactions of the bovine enzyme with NADPH. Published data, as well as new freeze-quench kinetic data presented here, are incompatible with the general opinion that NADPH and NADH react at the same site. Instead, it is proposed that these pyridine nucleotides react at opposite ends of the 90 Å long chain of prosthetic groups in Complex I. Ubiquinone is proposed to react with the Fe-S clusters in the TYKY subunit deep inside the hydrophilic domain. A new model for electron transfer in Complex I is proposed. In the accompanying paper this model is compared with the one advocated in current literature.
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Abdrakhmanova A, Zwicker K, Kerscher S, Zickermann V, Brandt U (2006) Tight binding of NADPH to the 39-kDa subunit of complex I is not required for catalytic activity but stabilizes the multiprotein complex. Biochim Biophys Acta 1757:1676–1682
Albracht SPJ (1974) Some new paramagnetic centers in submitochondrial particles detectable by EPR spectroscopy. Biochim Biophys Acta 347:183–192
Albracht SPJ (1980) The prosthetic groups in succinate dehydrogenase. Number and stoichiometry. Biochim Biophys Acta 612:11–28
Albracht SPJ (1982) In: Massey V, Williams CH (eds), Flavins and Flavoproteins: a possible solution to some unexplained observations on NADH dehydrogenase from beef-heart mitochondria. Elsevier/North-Holland Inc., Amsterdam, pp 759–762
Albracht SPJ (1984) In: Lee CP (eds), Current Topics in Bioenergetics: Applications of Electron Paramagnetic Resonance in the study of iron-sulfur clusters in energy-transducing membranes. Academic Press, New York, pp 79–106
Albracht SPJ (2010) The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited. II. Comparison of the proposed working hypothesis with literature data. J Bioener Biomem Accompanying paper. doi:10.1007/s10863-010-9302-y
Albracht SPJ, Subramanian J (1977) The number of Fe atoms in the iron-sulphur centers of the respiratory chain. Biochim Biophys Acta 462:36–48
Albracht SPJ, Bakker PT (1986) Evidence for two independent pathways of electron transfer in mitochondrial NADH:Q oxidoreductase. II. Kinetics of reoxidation of the reduced enzyme. Biochim Biophys Acta 850:423–428
Albracht SPJ, De Jong AMP (1997) Bovine-heart NADH:ubiquinone oxidoreductase is a monomer with 8 Fe-S clusters and 2 FMN groups. Biochim Biophys Acta 1318:92–106
Albracht SPJ, Hedderich R (2000) Learning from hydrogenases: location of a proton pump and of a second FMN in bovine NADH-ubiquinone oxidoreductase (Complex I). FEBS Lett 485:1–6
Albracht SPJ, Dooijewaard G, Leeuwerik FJ, Swol BV (1977) EPR signals of NADH: Q oxidoreductase. Shape and intensity. Biochim Biophys Acta 459:300–317
Albracht SPJ, Leeuwerik FJ, Van Swol B (1979) The stoichiometry of the iron-sulphur clusters 1a, 1b and 2 of NADH:Q oxidoreductase as present in beef-heart submitochondrial particles. FEBS Lett 104:197–200
Albracht SPJ, Van Verseveld HW, Hagen WR, Kalkman ML (1980) A comparison of the respiratory chain in particles from Paracoccus denitrificans and bovine heart mitochondria by EPR spectroscopy. Biochim Biophys Acta 593:173–186
Albracht SPJ, Van der Linden E, Faber BW (2003) Quantitative amino acid analysis of bovine NADH:ubiquinone oxidoreductase (Complex I) and related enzymes. Consequences for the number of prosthetic groups. Biochim Biophys Acta 1557:41–49
Bakker PT, Albracht SPJ (1986) Evidence for two independent pathways of electron transfer in mitochondrial NADH:Q oxidoreductase. I. Pre-steady-state kinetics with NADPH. Biochim Biophys Acta 850:413–422
Beechey RB, Holloway CT, Knight IG, Roberton AM (1966) Dicyclohexylcarbodiimide - An inhibitor of oxidative phosphorylation. Biochem Biophys Res Commun 23:75–80
Beechey RB, Roberton AM, Holloway CT, Knight IG (1967) The properties of dicyclohexylcarbodiimide as an inhibitor of oxidative phosphorylation. Biochemistry 6:3867–3879
Beinert H, Albracht SPJ (1982) New insights, ideas and unanswered questions concerning iron-sulfur clusters in mitochondria. Biochim Biophys Acta 683:245–277
Beinert H, Palmer G, Cremona T, Singer TP (1965) Kinetic studies on reduced diphosphopyridine nucleotide dehydrogenase by Electron Paramagnetic Resonance spectroscopy. J Biol Chem 240:475–480
Belogrudov G, Hatefi Y (1994) Catalytic sector of complex I (NADH:ubiquinone oxidoreductase): subunit stoichiometry and substrate-induced conformation changes. Biochemistry 33:4571–4576
Belogrudov GI, Hatefi Y (1996) Intersubunit interactions in the bovine mitochondrial complex I as revealed by ligand blotting. Biochem Biophys Res Commun 227:135–139
Berrisford JM, Sazanov LA (2009) Structural basis for the mechanism of respiratory complex I. J Biol Chem 284:29773–29783
Beyer RE, Brink TW, Crankshaw DL, Kuner JM, Pasternak A (1972) Effect of N, N′-dicyclohexylcarbodiimide and other carbodiimides on electron transfer catalyzed by submitochondrial particles. Biochemistry 11:961–969
Bridges HR, Grgic L, Harbour M, Hirst J (2009) The respiratory complexes I from the mitochondria of two Pichia species. Biochem J 422:151–159
Burgdorf T, Van der Linden E, Bernhard M, Yin QY, Back JW, Hartog AF, Muijsers AO, De Koster CG, Albracht SPJ, Friedrich B (2005) The soluble NAD+-reducing [NiFe]-hydrogenase from Ralstonia eutropha H16 consists of six subunits and can be specifically activated by NADPH. J Bacteriol 187:3122–3132
Carroll J, Fearnley IM, Skehel JM, Shannon RJ, Hirst J, Walker JE (2006) Bovine complex I is a complex of 45 different subunits. J Biol Chem 281:32724–32727
Chevallet M, Dupuis A, Issartel JP, Lunardi J, Van Belzen R, Albracht SPJ (2003) Two EPR-detectable [4Fe-4S] clusters, N2a and N2b, are bound to the NuoI (TYKY) subunit of NADH:ubiquinone oxidoreductase (Complex I) from Rhodobacter capsulatus. Biochim Biophys Acta 1557:51–66
Clason T, Ruiz T, Schägger H, Peng G, Zickermann V, Brandt U, Michel H, Radermacher M (2010) The structure of eukaryotic and prokaryotic complex I. J Struct Biol 169:81–88
Cleland KW, Slater EC (1953) Respiratory granules of heart muscle. Biochem J 53:547–556
Cremona T, Kearney EB (1964) Studies on the respiratory chain-linked NADH dehydrogenase. VI. Further purification and properties of the enzyme from beef heart. J Biol Chem 239:2328–2334
Darrouzet E, Issartel JP, Lunardi J, Dupuis A (1998) The 49-kDa subunit of NADH-ubiquinone oxidoreductase (Complex I) is involved in the binding of piericidin and rotenone, two quinone-related inhibitors. FEBS Lett 431:34–38
De Jong AMP, Albracht SPJ (1994) Ubisemiquinones as obligatory intermediates in the electron transfer from NADH to ubiquinone. Eur J Biochem 222:975–982
De Jong AMP, Kotlyar AB, Albracht SPJ (1994) Energy-induced structural changes in NADH:Q oxidoreductase of the mitochondrial respiratory chain. Biochim Biophys Acta 1186:163–171
Djafarzadeh R, Kerscher S, Zwicker K, Radermacher M, Lindahl M, Schägger H, Brandt U (2000) Biophysical and structural characterization of proton-translocating NADH-dehydrogenase (complex I) from the strictly aerobic yeast Yarrowia lipolytica. Biochim Biophys Acta 1459:230–238
Djavadi-Ohaniance L, Hatefi H (1975) Oxidation of NADPH by submitochondrial particles from beef heart in complete absence of transhydrogenase activity from NADPH to NAD. J Biol Chem 250:9397–9403
Dooijewaard G, Slater EC (1976) Steady-state kinetics of high molecular weight (type-I) NADH dehydrogenase. Biochim Biophys Acta 440:1–15
Earley FGP, Ragan CI (1984) Photoaffinity labelling of mitochondrial NADH dehydrogenase with arylazidoamorphigenin, an analogue of rotenone. Biochem J 224:525–534
Earley FGP, Patel SD, Ragan CI, Attardi G (1987) Photolabelling of a mitochondrially encoded subunit of NADH dehydrogenase with [3H]dihydrorotenone. FEBS Lett 219:108–113
Fearnley IM, Walker JE (1992) Conservation of sequences of subunits of mitochondrial complex I and their relationships with other proteins. Biochim Biophys Acta 1140:105–134
Finel M, Majander AS, Tyynelä J, De Jong AMP, Albracht SPJ, Wikström M (1994) Isolation and characterisation of subcomplexes of the mitochondrial NADH:ubiquinone oxidoreductase (complex I). Eur J Biochem 226:237–242
Fontecilla-Camps JC, Volbeda A, Cavazza C, Nicolet Y (2007) Structure/function relationships of [NiFe]- and [FeFe]-hydrogenases. Chem Rev 107:4273–4303
Friedrich T (2001) Complex I: a chimaera of a redox and conformation-driven proton pump? J Bioenerg Bioemembr 33:169–177
Galante YM, Hatefi Y (1979) Purification and molecular and enzymic properties of mitochondrial NADH dehydrogenase. Arch Biochem Biophys 192:559–568
Galkin AS, Grivennikova VG, Vinogradov AD (1999) H+/2e- stoichiometry in NADH-quinone reductase reactions catalyzed by bovine heart submitochondrial particles. FEBS Lett 451:157–161
Gornall AG, Bardawill CJ, David MM (1949) Deterermination of serum proteins by means of the biuret reaction. J Biol Chem 177:755–766
Grigorieff N (1998) Three-dimensional structure of bovine NADH:ubiquinone oxidoreductase (complex I) at 22 Å in ice. J Mol Biol 277:1033–1046
Guénebaut V, Vincentelli R, Mills D, Weiss H, Leonard KR (1997) Three-dimensional structure of NADH-dehydrogenase from Neurospora crassa by electron microscopy and conical tilt reconstruction. J Mol Biol 265:409–418
Guénebaut V, Schlitt A, Weiss H, Leonard K, Friedrich T (1998) Consistent structure between bacterial and mitochondrial NADH:ubiquinone oxidoreductase (Complex I). J Mol Biol 276:105–112
Gutman M, Singer TP, Casida JE (1970) Studies on the respiratory chain-linked reduced nicotinamide adenine dinucleotide dehydrogenase. XVII. Reaction sites of piericidin A and rotenone. J Biol Chem 245:1992–1997
Hatefi Y (1968) Flavoproteins of the electron transport system and the site of action of amytal, rotenone, and piericidin A. Proc Natl Acad Sci USA 60:733–740
Hatefi Y (1973) Oxidation of reduced triphosphopyridine nucelotide by sbmitochondrial particles from beef heart. Biochem Biophys Res Commun 50:978–984
Hatefi Y, Bearden AJ (1976) Electron paramagnetic resonance studies on the reduction of the components of complex I and transhydrogenase-inhibited complex I by NADH and NADPH. Biochem Biophys Res Commun 69:1032–1038
Hatefi Y, Hanstein WG (1973) Interactions of reduced and oxidized triphosphopyridine nucleotides with the electron-transport system of bovine heart mitochondria. Biochemistry 12:3515–3522
Hatefi Y, Stempel KE (1967) Resolution of complex I (DPNH-coenzyme Q reductase) of the mitochondrial electron transfer system. Biochem Biophys Res Commun 26:301–308
Hatefi Y, Stempel KE (1969) Isolation and enzymatic properties of the mitochondrial reduced diphosphopyridine nucleotide dehydrogenase. J Biol Chem 244:2350–2357
Hatefi Y, Yamaguchi M (1996) Nicotinamide nucleotide transhydrogenase: a model for utilization of substrate binding energy for proton translocation. FASEB J 10:444–452
Hatefi Y, Haavik AG, Griffiths DE (1962) Studies on the electron transfer system; XL. Preparation and properties of mitochondrial DPNH-Coenzyme Q reductase. J Biol Chem 237:1667–1680
Hearshen DO, Dunham WR, Albracht SPJ, Ohnishi T, Beinert H (1981) EPR spectral stimulation on cluster N-1b in NADH-ubiquinone oxidoreductase of bovine heart mitochondria. FEBS Lett 133:287–290
Hinchliffe P, Sazanov LA (2005) Organization of iron-sulfur clusters in respiratory Complex I. Science 309:771–774
Hirst J, Carroll J, Fearnley IM, Shannon RJ, Walker JE (2003) The nuclear encoded subunits of complex I from bovine heart mitochondria. Biochim Biophys Acta 1604:135–150
Hofhaus G, Weiss H, Leonard K (1991) Electron microscopic analysis of the peripheral and membrane parts of mitochondrial NADH dehydrogenase (complex I). J Mol Biol 221:1027–1043
Kotlyar AB, Vinogradov AD (1990) Slow active/inactive transition of the mitochondrial NADH-ubiquinone reductase. Biochim Biophys Acta 1019:151–158
Kowal AT, Morningstar JE, Johnson MK, Ramsay RR, Singer TP (1986) Spectroscopic characterization of the number and type of iron-sulfur clusters in NADH:ubiquinone oxidoreductase. J Biol Chem 261:9239–9245
Krishnamoorthy G, Hinkle PC (1988) Studies on the electron transfer pathway, topography of iron-sulfur centers, and site of coupling in NADH-Q oxidoreductase. J Biol Chem 263:17566–17575
Lenaz G (1998) Quinone specificity of Complex I. Biochim Biophys Acta 1364:207–221
Lenaz G, Daves GD Jr, Folkers K (1968) Organice structural specificity and sites of Coenzyme Q in succinoxidase and DPNH-oxidase systems. Arch Biochem Biophys 123:539–550
Lenaz G, Castelli A, Littarru GP, Bertoli E, Folkers K (1971) Specificity of lipids and Coenzyme Q in mitochondrial NADH and succin-oxidase of beef heart and S. cerevisiae. Arch Biochem Biophys 142:407–416
Leonard K, Haiker H, Weiss H (1987) Three-dimensional structure of NADH: ubiquinone reductase (complex I) from Neurospora mitochondria determined by electron microscopy of membrane crystals. J Mol Biol 194:277–286
Löw H, Vallin I (1963) Succinate-linked disphosphopyridine nucleotide reduction in submitochondrial particles. Biochim Biophys Acta 69:361–374
Lusty CJ, Machinist JM, Singer TP (1965) Studies on the respiratory chain-linked NADH dehydrogenase. VII; Labile sulfide groups in the dehydrogenase and in related proteins. J Biol Chem 240:1804–1810
Minakami S, Schindler FJ, Estabrook RW (1964) Hydrogen transfer between reduced diphosphopyridine nucleotide dehydrogenase and the respiratory chain. II. An initial lag in the oxidation of reduced diphosphopyridine nucleotide. J Biol Chem 239:2049–2054
Morgan DJ, Sazanov LA (2008) Three-dimensional structure of respiratory complex I from Escherichia coli in ice in the presence of nucleotides. Biochim Biophys Acta 1777:711–718
Murai M, Ishihara A, Nishioka T, Yagi T, Miyoshi H (2007) The ND1 subunit constructs the inhibitor binding domain in bovine heart mitochondrial complex I. Biochemistry 46:6409–6416
Ohnishi T (1975) Thermodynamic and EPR characterization of iron-sulfur centers in the NADH-ubiquinone segment of the mitochondrial respiratory chain in pigeon heart. Biochim Biophys Acta 387:475–490
Ohnishi T (1979) In: Capaldi RA (eds), Membrane Proteins in Energy Transduction: mitochondrial iron-sulfur flavodehydrogenases. Marcel Dekker, New York, pp 1–87
Ohnishi T (1998) Iron-sulfur clusters/semiquinones in Complex I. Biochim Biophys Acta 1364:186–206
Ohnishi T, Blum H, Galante YM, Hatefi Y (1981) Iron-sulfur N-1 clusters studied in NADH-ubiquinone oxidoreductase and in soluble NADH dehydrogenase. J Biol Chem 256:9216–9220
Orme-Johnson NR, Orme-Johnson WH, Hansen RE, Beinert H, Hatefi Y (1971) EPR detectable electron acceptors in submitochondrial particles from beef heart with specific reference to the iron-sulfur components of DPNH-ubiquinone reductase. Biochem Biophys Res Commun 44:446–452
Orme-Johnson NR, Hansen RE, Beinert H (1974a) Electron paramagnetic resonance-detectable electron acceptors in beef heart mitochondria. Reduced diphosphopyridine nucleotide ubiquinone reductase segment of the electron transfer system. J Biol Chem 249:1922–1927
Orme-Johnson NR, Hansen RE, Beinert H (1974b) Electron paramagnetic resonance-detectable electron acceptors in beef heart mitochondria. Ubihydroquinone-cytochrome c reductase segment of the electron transfer system and complex mitochondrial fragments. J Biol Chem 249:1928–1939
Paech C, Reynolds JR, Singer TP, Holm RH (1981) Structural identification of the iron-sulfur clusters of the respiratory chain-linked NADH dehydrogenase. J Biol Chem 256:3167–3170
Papa S, De Rasmo D, Scacco S, Signorile A, Technikova-Dobrova Z, Palmisano G, Sardanelli AM, Papa F, Panelli D, Scaringi R, Santeramo A (2008) Mammalian complex I: a regulable and vulnerable pacemaker in mitochondrial respiratory function. Biochim Biophys Acta 1777:719–728
Peng G, Fritzsch G, Zickermann V, Schagger H, Mentele R, Lottspeich F, Bostina M, Radermacher M, Huber R, Stetter KO, Michel H (2003) Isolation, characterization and electron microscopic single particle analysis of the NADH:ubiquinone oxidoreductase (complex I) from the hyperthermophilic eubacterium Aquifex aeolicus. Biochemistry 42:3032–3039
Pilkington SJ, Skehel JM, Gennis RB, Walker JE (1991) Relationship between mitochondrial NADH-ubiquinone reductase and a bacterial NAD-reducing hydrogenase. Biochemistry 30:2166–2175
Prieur I, Lunardi J, Dupuis A (2001) Evidence for a quinone binding site close to the interface between NUOD and NUOB subunits of Complex I. Biochim Biophys Acta 1504:173–178
Radermacher M, Ruiz T, Clason T, Benjamin S, Brandt U, Zickermann V (2006) The three-dimensional structure of complex I from Yarrowia lipolytica: a highly dynamic enzyme. J Struct Biol 154:269–279
Ragan CI (1976) The interaction of reduced nicotinamide-adenine dinucleotide phosphate with reduced nicotinamide-adenine dinucleotide-ubiquinone reductase from bovine heart mitochondria. Biochem J 158:149–151
Ragan CI, Widger WR, King TE (1974) Pyridine nucleotide transhydrogenase activity of soluble cardiac NADH dehydrogenase and particulate NADH-ubiquinone reductase. Biochem Biophys Res Commun 60:894–900
Reda T, Barker CD, Hirst J (2008) Reduction of the iron sulfur clusters in mitochondrial NADH:ubiquinone oxidoreductase (complex I) by EuII-DTPA, a very low potential reductant. Biochemistry 47:8885–8893
Ringler RL, Minakami S, Singer TP (1960) Isolation and properties of the DPNH dehydrogenase of the respiratory chain from heart mitochondria. Biochem Biophys Res Commun 3:417–422
Ringler RL, Minakami S, Singer TP (1963) Studies on the respiratory chain-linked NADH dehydrogenase. II. Isolation and molecular properties of the enzyme from beef heart. J Biol Chem 238:801–810
Rydström J, Teixeira da Cruz A, Ernster L (1971) Steady-state kinetics of mitochondrial nicotinamide nucleotide transhydrogenase. 2. The energy-linked reaction. Eur J Biochem 23:213–219
Rydström J, Montelius J, Bäckström D, Ernster L (1978) The mechanism of oxidation of reduced nicotinamide dinucleotide phosphate by submitochondrial particles from beef heart. Biochim Biophys Acta 501:370–380
Sazanov LA, Hinchliffe P (2006) Structure of the hydrophilic domain of respiratory Complex I from Thermus thermophilus. Science 311:1430–1436
Schulte U, Haupt V, Abelmann A, Fecke W, Brors B, Rasmussen T, Friedrich T, Weiss H (1999) A reductase/isomerase subunit of mitochondrial NADH:ubiquinone oxidoreductase (complex I) carries an NADPH and is involved in the biogenesis of the complex. J Mol Biol 292:569–580
Singer TP (1979) Mitochondrial electron-transport inhibitors. Meth Enzymol 55:454–462
Van Belzen R (1991) Electron transfer in NADH:Q oxidoreductase from bovine-heart mitochondria. Ph.D. Thesis, University of Amsterdam, Amsterdam, The Netherlands
Van Belzen R, Albracht SPJ (1989) The pathway of electron transfer in NADH:Q oxidoreductase. Biochim Biophys Acta 974:311–320
Van Belzen R, Van Gaalen MC, Cuypers PA, Albracht SPJ (1990) New evidence for the dimeric nature of NADH:Q oxidoreductase in bovine-heart submitochondrial particles. Biochim Biophys Acta 1017:152–159
Van Belzen R, De Jong AMP, Albracht SPJ (1992) On the stoichiometry of the iron-sulphur clusters in mitochondrial NADH:ubiquinone oxidoreductase. Eur J Biochem 209:1019–1022
Van Belzen R, Kotlyar AB, Moon N, Dunham WR, Albracht SPJ (1997) The iron-sulfur clusters 2 and ubisemiquinone radicals of NADH:ubiquinone oxidoreductase are involved in energy coupling in submitochondrial particles. Biochemistry 36:886–893
Van der Linden E, Faber BW, Bleijlevens B, Burgdorf T, Bernhard M, Friedrich B, Albracht SPJ (2004) Selective release and function of one of the two FMN groups in the cytoplasmic NAD+-reducing [NiFe]-hydrogenase from Ralstonia eutropha. Eur J Biochem 271:801–808
Vanneste WH (1966) Molecular proportion of the fixed cytochrome components of the respiratory chain of Keilin-Hartree particles and beef heart mitochondria. Biochim Biophys Acta 113:175–178
Volbeda A, Charon MH, Piras C, Hatchikian EC, Frey M, Fontecilla-Camps JC (1995) Crystal structure of the nickel-iron hydrogenase from Desulfovibrio gigas. Nature 373:580–587
Volbeda A, Garcia E, Piras C, De Lacey AL, Fernandez VM, Hatchikian EC, Frey M, Fontecilla-Camps JC (1996) Structure of the [NiFe] hydrogenase active site: Evidence for biologically uncommon Fe ligands. J Am Chem Soc 118:12989–12996
Walker JE (1992) The NADH:ubiquinone oxidoreductase (complex I) of respiratory chains. Q Rev Biophys 25:253–324
Walker JE, Arizmendi JM, Dupuis A, Fearnley IM, Finel M, Medd SM, Pilkington SJ, Runswick MJ, Skehel JM (1992) Sequences of 20 subunits of NADH:ubiquinone oxidoreductase from bovine heart mitochondria. Application of a novel strategy for sequencing proteins using the polymerase chain reaction. J Mol Biol 226:1051–1072
Wikström M (1984) Two protons are pumped from the mitochondrial matrix per electron transferred between NADH and ubiquinone. FEBS Lett 169:300–304
Yagi T (1987) Inhibition of NADH-ubiquinone reductase activity by N, N′- dicyclohexylcarbodiimide and correlation of this inhibition with the occurrence of energy-coupling site 1 in various organisms. Biochemistry 26:2822–2828
Yagi T, Hatefi Y (1988) Identification of the dicyclohexylcarbodiimide-binding subunit of NADH- ubiquinone oxidoreductase (Complex I). J Biol Chem 263:16150–16155
Yagi T, Matsuno-Yagi A (2003) The Proton-Translocating NADH-Quinone Oxidoreductase in the Respiratory Chain: The Secret Unlocked. Biochemistry 42:2266–2274
Yakovlev G, Reda T, Hirst J (2007) Reevaluating the relationship between EPR spectra and enzyme structure for the iron–sulfur clusters in NADH:quinone oxidoreductase. Proc Natl Acad Sci USA 104:1270–12725
Yamaguchi M, Belogrudov GI, Hatefi Y (1998) Mitochondrial NADH-ubiquinone oxidoreductase (Complex I). Effect of substrates on the fragmentation of subunits by trypsin. J Biol Chem 273:8094–8098
Yamaguchi M, Belogrudov GI, Matsuno-Yagi A, Hatefi Y (2000) The multiple nicotinamide nucleotide-binding subunits of bovine heart mitochondrial NADH:ubiquinone oxidoreductase (complex I). Eur J Biochem 267:329–336
Yano T, Sklar J, Nakamaru-Ogiso E, Takahashi Y, Yagi T, Ohnishi T (2003) Characterization of cluster N5 as a fast-relaxing [4Fe-4S] cluster in the Nqo3 subunit of the proton-translocating NADH-ubiquinone oxidoreductase from Paracoccus denitrificans. J Biol Chem 278:15514–15522
Yano T, Dunham WH, Ohnishi T (2005) Characterization of the Δμ H+-sensitive ubisemiquinone species (SQNf) and the interaction with cluster N2: new insight into the energy-coupled electron transfer in Complex I. Biochemistry 44:1744–1754
Zickermann V, Kerscher S, Zwicker K, Tocilescu MA, Radermacher M, Brandt U (2009) Architecture of complex I and its implications for electron transfer and proton pumping. Biochim Biophys Acta 1787:574–583
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Albracht, S.P.J. The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited. J Bioenerg Biomembr 42, 261–278 (2010). https://doi.org/10.1007/s10863-010-9301-z
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DOI: https://doi.org/10.1007/s10863-010-9301-z