McDonald 2009 FEBS Lett
| McDonald BM, Wydrob MM, Lightowlers RN, Lakey JH (2009) Probing the orientation of yeast VDAC1 in vivo. FEBS Lett 583:739-42. |
McDonald BM, Wydrob MM, Lightowlers RN, Lakey JH (2009) FEBS Lett
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 beta-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. • Keywords: Mitochondrial membrane channels
Labels: MiParea: Respiration, Genetic knockout;overexpression
Organism: Saccharomyces cerevisiae
Preparation: Intact cells Enzyme: Inner mt-membrane transporter
HRR: Oxygraph-2k
