Probing the structure of the mitochondrial channel,VDAC, by site-directed mutagenesis: A progress report |
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Authors: | Elizabeth Blachly-Dyson Song Zhi Peng Marco Colombini Michael Forte |
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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 |
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Abstract: | The voltage-dependent anion-selective channel (VDAC) of the mitochondrial outer membrane is formed by a small ( 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. |
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Keywords: | Yeast VDAC oligonucleotide-directed mutagenesis ion selectivity voltage gating mitochondrial outer membrane anion channel ion channel |
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