The Respiratory Complex I (NDH I) from Klebsiella pneumoniae, a Sodium Pump

Gemperli, Anja C.; Dimroth, Peter; Steuber, Julia

doi:10.1074/jbc.m204860200

Cited by 62 publications

(84 citation statements)

References 53 publications

(63 reference statements)

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“…Over the last years, we accumulated evidence that NDH I from enterobacteria like Klebsiella pneumoniae (Gemperli et al 2002(Gemperli et al , 2003 or E. coli (Steuber et al 2000) translocates Na + rather than protons. Our views have been challenged by the hypothesis that complex I from K. pneumoniae (Bertsova and Bogachev 2004) or E. coli (Stolpe and Friedrich 2004) exclusively acts as a proton pump.…”

Section: Discussionmentioning

confidence: 99%

“…Here, Na + translocation was strictly coupled to the oxidation of NADH even in the presence of a chemical Na + gradient which would have favored uptake of Na + by the proteoliposomes (Steuber et al 2000;Gemperli et al 2002Gemperli et al , 2003. Obviously, Na + transport through the NuoL subunit in the holo-NDH I is prevented, most likely by the interaction of NuoL with other components of the complex.…”

Section: Discussionmentioning

confidence: 99%

“…The vesicles were collected by a second centrifugation (20,000£g, 30 min), resuspended in 1-1.5 ml buVer (25-32 mg protein ml ¡1 ), shock frozen in liquid nitrogen and stored at ¡80°C until use. Na + and K + transport Uptake of Na + by ER membrane vesicles was followed by atomic absorption spectroscopy (Gemperli et al 2002), modiWed as follows. Reaction mixtures contained in 0.4 ml at room temperature: ER vesicles (2 mg protein), 10 mM Tris/MES, pH 8.0, and as denoted 5 or10 mM choline chloride and 5 or 10 mM KCl.…”

Section: Expression Of Nuol N and Fractionation Of Membranesmentioning

confidence: 99%

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Transport of Na+ and K+ by an antiporter-related subunit from the Escherichia coli NADH dehydrogenase I produced in Saccharomyces cerevisiae

et al. 2007

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The NADH dehydrogenase I from Escherichia coli is a bacterial homolog of the mitochondrial complex I which translocates Na + rather than H + . To elucidate the mechanism of Na + transport, the C-terminally truncated NuoL subunit (NuoL N ) which is related to Na + /H + antiporters was expressed as a protein A fusion protein (ProtANuoL N ) in the yeast Saccharomyces cerevisiae which lacks an endogenous complex I. The fusion protein inserted into membranes from the endoplasmatic reticulum (ER), as conWrmed by diVerential centrifugation and Western analysis. Membrane vesicles containing ProtA-NuoL N catalyzed the uptake of Na + and K + at rates which were signiWcantly higher than uptake by the control vesicles under identical conditions, demonstrating that ProtA-NuoL N translocated Na + and K + independently from other complex I subunits. Na + transport by ProtA-NuoL N was inhibited by EIPA (5-(N-ethyl-N-isopropyl)-amiloride) which speciWcally reacts with Na + /H + antiporters. The cation selectivity and function of the NuoL subunit as a transporter module of the NADH dehydrogenase complex is discussed.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Expression Of Nuol N and Fractionation Of Membranesmentioning

confidence: 99%

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Transport of Na+ and K+ by an antiporter-related subunit from the Escherichia coli NADH dehydrogenase I produced in Saccharomyces cerevisiae

et al. 2007

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“…The data suggested that inverted membrane vesicles from E. coli catalyzed NADH-driven Na 1 uptake, which was prevented by inhibitors of complex I but not by protonophores (22). Complexes I from K. pneumoniae and Y. lipolytica, reconstituted into liposomes, were reported to act as Na 1 pumps with a Na 1 /e stoichiometry of 1 (21,23). These observations were contested by other authors that have shown that complexes I from E. coli and K. pneumoniae pump H 1 (24,25).…”

Section: Na 1 As the Coupling Ionmentioning

confidence: 92%

“…Steuber and coworkers (20,21) proposed that some complexes I use Na 1 instead of the consensual H 1 . Transport of The enzyme is an L-shaped membrane protein, consisting of a peripheral and a membrane arm.…”

Section: Na 1 As the Coupling Ionmentioning

confidence: 99%

The role of proton and sodium ions in energy transduction by respiratory complex I

2012

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SummaryRespiratory complex I plays a central role in energy transduction. It catalyzes the oxidation of NADH and the reduction of quinone, coupled to cation translocation across the membrane, thereby establishing an electrochemical potential. For more than half a century, data on complex I has been gathered, including recently determined crystal structures, yet complex I is the least understood complex of the respiratory chain. The mechanisms of quinone reduction, charge translocation and their coupling are still unknown. The H 1 is accepted to be the coupling ion of the system; however, Na 1 has also been suggested to perform such a role. In this article, we address the relation of those two ions with complex I and refer ion pump and Na 1 /H 1 antiporter as possible transport mechanisms of the system. We put forward a hypothesis to explain some apparently contradictory data on the nature of the coupling ion, and we revisit the role of H 1 and Na 1 cycles in the overall bioenergetics of the cell. IUBMBIUBMB Life, 64(6): 492-498, 2012

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Dicyclohexyl Carbodiimide

2007

Hawley's Condensed Chemical Dictionary

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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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The Respiratory Complex I (NDH I) from Klebsiella pneumoniae, a Sodium Pump

Abstract: The electrogenic NADH:Q oxidoreductase from the enterobacterium Klebsiella pneumoniae transports Na ؉ ions. The complex was purified with an increase of the specific Na ؉ transport activity from 0.

Cited by 62 publications

References 53 publications

Transport of Na+ and K+ by an antiporter-related subunit from the Escherichia coli NADH dehydrogenase I produced in Saccharomyces cerevisiae

Transport of Na+ and K+ by an antiporter-related subunit from the Escherichia coli NADH dehydrogenase I produced in Saccharomyces cerevisiae

The role of proton and sodium ions in energy transduction by respiratory complex I

Dicyclohexyl Carbodiimide

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