Recombinant yeast VDAC2: a comparison of electrophysiological features with the native form

FEBS Open Bio. 2019 Jul;9(7):1184-1193. doi: 10.1002/2211-5463.12574. Epub 2019 Jun 17.

Abstract

Voltage-dependent anion channel isoform 2 of the yeast Saccharomyces cerevisiae (yVDAC2) was believed for many years to be devoid of channel activity. Recently, we isolated yVDAC2 and showed that it exhibits channel-forming activity in the planar lipid bilayer system when in its so-called native form. Here, we describe an alternative strategy for yVDAC2 isolation, through heterologous expression in bacteria and refolding in vitro. Recombinant yVDAC2, like its native form, is able to form voltage-dependent channels. However, some differences between native and recombinant yVDAC2 emerged in terms of voltage dependence and ion selectivity, suggesting that, in this specific case, the recombinant protein might be depleted of post-translational modification(s) that occur in eukaryotic cells.

Keywords: VDAC2; electrophysiology; heterologous expression; mitochondria; mitochondrial porins; planar lipid bilayer; yVDAC2; yeast; yeastVDAC2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Electrophysiological Phenomena
  • Intracellular Membranes / metabolism
  • Lipid Bilayers / metabolism
  • Mitochondria / metabolism
  • Protein Engineering / methods*
  • Protein Isoforms / metabolism
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / metabolism
  • Voltage-Dependent Anion Channel 2 / metabolism*
  • Voltage-Dependent Anion Channel 2 / physiology*
  • Voltage-Dependent Anion Channels / metabolism

Substances

  • Lipid Bilayers
  • Protein Isoforms
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins
  • Voltage-Dependent Anion Channel 2
  • Voltage-Dependent Anion Channels