Inhibition of NADH-ubiquinone reductase by N,N'-dicyclohexylcarbodiimide and correlation of this inhibition with the occurrence of energy-coupling site 1 in various organisms

Yagi, Takao

doi:10.1021/bi00384a025

Cited by 79 publications

(52 citation statements)

References 52 publications

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“…Once unlocked, this oxidation is inhibited by rotenone or DCCD. The inhibition by DCCD is consistent with a direct coupling between electron transfer and proton translocation as earlier suggested for the oxidation of NADH (Yagi 1987;Yagi and Hatefi 1988). Rotenone and piericidin A can each bind to two sites, one close to a site in the 49-kDa subunit, the site structurally equivalent to that of the Ni-Fe site in [NiFe]-hydrogenases (Darrouzet et al 1998;Prieur et al 2001), and the other on Table 2 Kinetics of the reactions of NADH and NADPH with bovine Complex I-The NAD(P)H→FMN data are from freeze-quench experiments.…”

Section: Kinetic Issues Reactions With Nad(p)hsupporting

confidence: 89%

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The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited

Albracht

2010

J Bioenerg Biomembr

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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 longknown 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 90Å 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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Section: Kinetic Issues Reactions With Nad(p)hsupporting

confidence: 89%

“…It has been reported that DCCD binds mainly to the ND1 subunit in Hatefi's Complex I (Yagi 1987;Yagi and Hatefi 1988). The labelling was not affected by the complete inhibition by rotenone, although the ND1 subunit can bind this inhibitor as well (Earley and Ragan 1984;Earley et al 1987).…”

Section: >200 Msmentioning

confidence: 88%

The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited

Albracht

2010

J Bioenerg Biomembr

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“…4). Subunit 1 has been postulated to be a transmembrane protein and a site of proton translocation and energy coupling (59,60). In addition, subunit 1 has been proposed as the site for quinone binding (23), although an isolated subcomplex of bovine complex I in which ND-1 was absent was able to couple NADH oxidation to quinone reduction (20).…”

Section: Resultsmentioning

confidence: 99%

Characterization of the region encoding the CO-induced hydrogenase of Rhodospirillum rubrum

et al. 1996

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In the photosynthetic bacterium Rhodospirillum rubrum, the presence of carbon monoxide (CO) induces expression of several proteins. These include carbon monoxide dehydrogenase (CODH) and a CO-tolerant hydrogenase. Together these enzymes catalyze the following conversion: CO ؉ H 2 O 3 CO 2 ؉ H 2 . This system enables R. rubrum to grow in the dark on CO as the sole energy source. Expression of this system has been shown previously to be regulated at the transcriptional level by CO. We have now identified the remainder of the CO-regulated genes encoded in a contiguous region of the R. rubrum genome. These genes, cooMKLXU, apparently encode proteins related to the function of the CO-induced hydrogenase. As seen before with the gene for the large subunit of the CO-induced hydrogenase (cooH), most of the proteins predicted by these additional genes show significant sequence similarity to subunits of Escherichia coli hydrogenase 3. In addition, all of the newly identified coo gene products show similarity to subunits of NADH-quinone oxidoreductase (energyconserving NADH dehydrogenase I) from various eukaryotic and prokaryotic organisms. We have found that dicyclohexylcarbodiimide, an inhibitor of mitochondrial NADH dehydrogenase I (also called complex I), inhibits the CO-induced hydrogenase as well. We also show that expression of the cooMKLXUH operon is regulated by CO and the transcriptional activator CooA in a manner similar to that of the cooFSCTJ operon that encodes the subunits of CODH and related proteins.In the presence of carbon monoxide (CO), Rhodospirillum rubrum induces synthesis of a CO-oxidizing system. This system catalyzes the net reaction CO ϩ H 2 O 3 CO 2 ϩ H 2 . The reaction is carried out by two enzymes: CO dehydrogenase (CODH) and a CO-tolerant hydrogenase. CODH is a wellcharacterized nickel-iron enzyme (10) that carries out the oxidation of CO to CO 2 , producing two reducing equivalents. Hydrogenase then consumes these reducing equivalents by the reduction of two protons to H 2 (9, 17). Intermediate electron carriers (including the ferredoxin-like small subunit of CODH [17]) may also be involved in the reaction.The hydrogenase is tightly membrane bound and has yet to be purified, although the sequence of its large subunit (CooH) and several of its properties have been described previously (22). A wide variety of hydrogenases from other organisms, however, have been purified and characterized. These hydrogenases have been found to fall into three distinct categories: Fe-only, Ni-Fe, and Ni-Fe-Se enzymes (for reviews, see references 1, 4, and 56). The CO-induced hydrogenase from R. rubrum apparently belongs to the Ni-Fe class, on the basis of its protein sequence and the requirement of Ni for activity (22).There seem to be two CO-regulated transcripts in R. rubrum. The first, cooFSCTJ, encodes CODH and related proteins (27,29,43) and has been shown to be regulated by the product of the cooA gene (43). CooA appears to bind to a specific region of DNA upstream of the cooF promoter in a CO-dependent m...

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“…The carboxyl group-modifying reagent DCCD specifically inhibits mitochondrial complex I and the related NDH-1 from bacteria (51). In bovine complex I, DCCD modifies the membrane-spanning ND1 subunit (NuoH homologue in NDH-1) and inhibits both electron transfer and proton transport (52).…”

Section: Vol 188 2006 Sodium Ion Binding To Complex I From K Pneummentioning

confidence: 99%

Specific Modification of a Na ⁺ Binding Site in NADH:Quinone Oxidoreductase from Klebsiella pneumoniae with Dicyclohexylcarbodiimide

2006

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The respiratory NADH:quinone oxidoreductase (complex I) (NDH-1) is a multisubunit enzyme that translocates protons (or in some cases Na ؉ ) across energy-conserving membranes from bacteria or mitochondria. We studied the reaction of the Na ؉ -translocating complex I from the enterobacterium Klebsiella pneumoniae with N,N-dicyclohexylcarbodiimide (DCCD), with the aim of identifying a subunit critical for Na ؉ binding. At low Na ؉ concentrations (0.6 mM), DCCD inhibited both quinone reduction and Na ؉ transport by NDH-1 concurrent with the covalent modification of a 30-kDa polypeptide. In the presence of 50 mM Na ؉ , NDH-1 was protected from inhibition by DCCD, and the modification of the 30-kDa polypeptide with [ 14 C]DCCD was prevented, indicating that Na ؉ and DCCD competed for the binding to a critical carboxyl group in NDH-1. The 30-kDa polypeptide was assigned to NuoH, the homologue of the ND1 subunit from mitochondrial complex I. It is proposed that Na ؉ binds to the NuoH subunit during NADH-driven Na ؉ transport by NDH-1.

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Inhibition of NADH-ubiquinone reductase by N,N'-dicyclohexylcarbodiimide and correlation of this inhibition with the occurrence of energy-coupling site 1 in various organisms

Cited by 79 publications

References 52 publications

The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited

The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited

Characterization of the region encoding the CO-induced hydrogenase of Rhodospirillum rubrum

Specific Modification of a Na ⁺ Binding Site in NADH:Quinone Oxidoreductase from Klebsiella pneumoniae with Dicyclohexylcarbodiimide

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Inhibition of NADH-ubiquinone reductase by N,N'-dicyclohexylcarbodiimide and correlation of this inhibition with the occurrence of energy-coupling site 1 in various organisms

Cited by 79 publications

References 52 publications

The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited

The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited

Characterization of the region encoding the CO-induced hydrogenase of Rhodospirillum rubrum

Specific Modification of a Na + Binding Site in NADH:Quinone Oxidoreductase from Klebsiella pneumoniae with Dicyclohexylcarbodiimide

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Specific Modification of a Na ⁺ Binding Site in NADH:Quinone Oxidoreductase from Klebsiella pneumoniae with Dicyclohexylcarbodiimide