FEBS Letters
Volume 583, Issue 4 , Pages 739-742, 18 February 2009

Probing the orientation of yeast VDAC1 in vivo

Edited by Peter Brzezinski

  • Beth M. McDonald

      Affiliations

    • The Institute for Cell and Molecular Biosciences, Newcastle University, Framlington Place, Newcastle-upon-Tyne NE2 4HH, UK
    • Mitochondrial Research Group, Newcastle University, Framlington Place, Newcastle-upon-Tyne NE2 4HH, UK
    • ,
  • Mateusz M. Wydro

      Affiliations

    • Mitochondrial Research Group, Newcastle University, Framlington Place, Newcastle-upon-Tyne NE2 4HH, UK
    • ,
  • Robert N. Lightowlers

      Affiliations

    • Mitochondrial Research Group, Newcastle University, Framlington Place, Newcastle-upon-Tyne NE2 4HH, UK
    • ,
  • Jeremy H. Lakey

      Affiliations

    • The Institute for Cell and Molecular Biosciences, Newcastle University, Framlington Place, Newcastle-upon-Tyne NE2 4HH, UK
    • Corresponding Author InformationCorresponding author. Fax: +44 191 222 7424.

Received 11 December 2008; received in revised form 13 January 2009; accepted 14 January 2009. published online 28 January 2009.

Abstract 

Voltage dependent anion channel (VDAC) is a vital ion channel in mitochondrial outer membranes and its structure was recently shown to be a 19 stranded β-barrel. However the orientation of VDAC in the membrane remains unclear. We probe here the topology and membrane orientation of yeast Saccharomyces cerevisiae in vivo. Five FLAG-epitopes were independently inserted into scVDAC1 and their surface exposure in intact and disrupted mitochondria detected by immunoprecipitation. Functionality was confirmed by measurements of respiration. Two epitopes suggest that VDAC (scVDAC) has its C-terminus exposed to the cytoplasm whilst two others are more equivocal and, when combined with published data, suggest a dynamic behavior.

Abbreviations: OMM, outer mitochondrial membrane, IMS, intermembrane space, scVDAC, Saccharomyces cerevisiae VDAC, hsVDAC, human VDAC, IP, immunoprecipitation

Keywords: Voltage dependent anion channel, Mitochondrial porin, Structure, Immunoprecipitation, Yeast

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PII: S0014-5793(09)00045-3

doi:10.1016/j.febslet.2009.01.039

FEBS Letters
Volume 583, Issue 4 , Pages 739-742, 18 February 2009

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