The supramolecular assemblies of voltage-dependent anion channels in the native membrane.
Voltage-dependent anion channels (VDACs) are major constituents of the outer mitochondrial membrane (OMM). These primary transporters of nucleotides, ions and metabolites mediate a substantial portion of the OMM molecular traffic. To study the native supramolecular organization of the VDAC, we have isolated, characterized and imaged OMMs from potato tubers. SDS-PAGE and mass spectrometry of OMMs revealed the presence of the VDAC isoforms POM34 and POM36, as well as the translocase of the OMM complex. Tubular two-dimensional crystals of the VDAC spontaneously formed after incubation of OMMs for two to three months at 4 degrees C. Transmission electron microscopy revealed an oblique lattice and unit cells housing six circular depressions arranged in a hexagon. Atomic force microscopy of freshly isolated OMMs demonstrated (i) the existence of monomers to tetramers, hexamers and higher oligomers of the VDAC and (ii) its spatial arrangement within the oligomers in the native membrane. We discuss the importance of the observed oligomerization for modulation of the VDAC function, for the binding of hexokinase and creatine kinase to the OMM and for mitochondria-mediated apoptosis.
Hoogenboom
Bart W
BW
M. E. Müller Institute for Microscopy, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland.
Suda
Kitaru
K
Engel
Andreas
A
Fotiadis
Dimitrios
D
D017209:Apoptosis; D003402:Creatine Kinase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006593:Hexokinase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D018625:Microscopy, Atomic Force; D046529:Microscopy, Electron, Transmission; D051336:Mitochondrial Membranes / Q000737:chemistry* / Q000648:ultrastructure; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011198:Solanum tuberosum / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D050994:Voltage-Dependent Anion Channels / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D010940:Plant Proteins; D020033:Protein Isoforms; D050994:Voltage-Dependent Anion Channels; D006593:Hexokinase; D003402:Creatine Kinase