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Probing the structure of the mitochondrial channel,VDAC, by site-directed mutagenesis: A progress report
Authors:Elizabeth Blachly-Dyson  Song Zhi Peng  Marco Colombini  Michael Forte
Institution:(1) Vollum Institute for Advanced Biomedical Research, Oregon Health Sciences University, 97201 Portland, Oregon;(2) Department of Zoology, University of Maryland, 20742 College Park, Maryland
Abstract:The voltage-dependent anion-selective channel (VDAC) of the mitochondrial outer membrane is formed by a small (sim 30 kDa) polypeptide, but shares with more complex channels the properties of voltage-dependent gating and ion selectivity. Thus, it is a useful model for studying these properties. The molecular biology techniques available in yeast allow us to construct mutant versions of the cloned yeast VDAC genein vitro, using oligonucleotide-directed mutagenesis, and to express the mutant genes in yeast cells in the absence of wild-type VDAC. We find that one substitution mutation (lys 61 to glu) alters the selectivity of VDAC.
Keywords:Yeast VDAC  oligonucleotide-directed mutagenesis  ion selectivity  voltage gating  mitochondrial outer membrane  anion channel  ion channel
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