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Biochemistry (Moscow)

, Volume 73, Issue 2, pp 123–129 | Cite as

Site-directed mutagenesis of conserved cysteine residues in NqrD and NqrE subunits of Na+-translocating NADH:quinone oxidoreductase

  • M. S. Fadeeva
  • Y. V. Bertsova
  • M. I. Verkhovsky
  • A. V. BogachevEmail author
Accelerated Publication

Abstract

Each of two hydrophobic subunits of Na+-translocating NADH:quinone oxidoreductase (NQR), NqrD and NqrE, contain a pair of strictly conserved cysteine residues within their transmembrane α-helices. Site-directed mutagenesis showed that substitutions of these residues in NQR of Vibrio harveyi blocked the Na+-dependent and 2-n-heptyl-4-hydroxyquinoline N-oxide-sensitive quinone reductase activity of the enzyme. However, these mutations did not affect the interaction of NQR with NADH and menadione. It was demonstrated that these conserved cysteine residues are necessary for the correct folding and/or the stability of the NQR complex. Mass and EPR spectroscopy showed that NQR from V. harveyi bears only a 2Fe-2S cluster as a metal-containing prosthetic group.

Key words

Na+-translocating NADH:quinone oxidoreductase sodium potential Vibrio respiratory chain 

Abbreviations

Ap

ampicillin

dNADH

reduced nicotinamide hypoxanthine dinucleotide

Em

midpoint redox potential

EPR

electronic paramagnetic resonance

HQNO

2-n-heptyl-4-hydroxyquinoline N-oxide

K3

menadione

NQR

Na+-translocating NADH:quinone oxidoreductase

Rf

rifampicin

SBP

sub-bacterial particles

Sm

streptomycin

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Copyright information

© Pleiades Publishing, Ltd. 2008

Authors and Affiliations

  • M. S. Fadeeva
    • 1
  • Y. V. Bertsova
    • 2
  • M. I. Verkhovsky
    • 3
  • A. V. Bogachev
    • 2
    Email author
  1. 1.Department of Bioinformatics and BioengineeringLomonosov Moscow State UniversityMoscowRussia
  2. 2.Department of Molecular Energetics of Microorganisms, Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia
  3. 3.Helsinki Bioenergetics Group, Institute of BiotechnologyUniversity of HelsinkiHelsinkiFinland

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