Locking loop movement in the ubiquinone pocket of complex I disengages the proton pumps

Nat Commun. 2018 Oct 29;9(1):4500. doi: 10.1038/s41467-018-06955-y.


Complex I (proton-pumping NADH:ubiquinone oxidoreductase) is the largest enzyme of the mitochondrial respiratory chain and a significant source of reactive oxygen species (ROS). We hypothesized that during energy conversion by complex I, electron transfer onto ubiquinone triggers the concerted rearrangement of three protein loops of subunits ND1, ND3, and 49-kDa thereby generating the power-stoke driving proton pumping. Here we show that fixing loop TMH1-2ND3 to the nearby subunit PSST via a disulfide bridge introduced by site-directed mutagenesis reversibly disengages proton pumping without impairing ubiquinone reduction, inhibitor binding or the Active/Deactive transition. The X-ray structure of mutant complex I indicates that the disulfide bridge immobilizes but does not displace the tip of loop TMH1-2ND3. We conclude that movement of loop TMH1-2ND3 located at the ubiquinone-binding pocket is required to drive proton pumping corroborating one of the central predictions of our model for the mechanism of energy conversion by complex I proposed earlier.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography, X-Ray
  • Disulfides
  • Electron Transport
  • Electron Transport Complex I / chemistry*
  • Electron Transport Complex I / genetics
  • Electron Transport Complex I / ultrastructure*
  • Enzyme Activation
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / genetics
  • Kinetics
  • Mitochondrial Membranes / enzymology
  • Mitochondrial Membranes / metabolism
  • Models, Molecular
  • Molecular Dynamics Simulation
  • Mutagenesis, Site-Directed
  • Protein Conformation
  • Proton Pumps / chemistry*
  • Proton Pumps / ultrastructure
  • Reactive Oxygen Species / metabolism
  • Ubiquinone / chemistry*
  • Ubiquinone / ultrastructure*
  • Yarrowia / genetics
  • Yarrowia / metabolism


  • Disulfides
  • Escherichia coli Proteins
  • Proton Pumps
  • Reactive Oxygen Species
  • Ubiquinone
  • Electron Transport Complex I