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Probing the orientation of yeast VDAC1 in vivo

Lookup NU author(s): Dr Mateusz Wydro, Professor Robert Lightowlers, Professor Jeremy LakeyORCiD

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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 (scV-DAC) has its C-terminus exposed to the cytoplasm whilst two others are more equivocal and, when combined with published data, suggest a dynamic behavior. (C) 2009 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Publication metadata

Author(s): McDonald BM, Wydro MM, Lightowlers RN, Lakey JH

Publication type: Article

Publication status: Published

Journal: FEBS Letters

Year: 2009

Volume: 583

Issue: 4

Pages: 739-742

ISSN (print): 0014-5793

ISSN (electronic): 1873-3468

Publisher: Elsevier BV

URL: http://dx.doi.org/10.1016/j.febslet.2009.01.039

DOI: 10.1016/j.febslet.2009.01.039

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