title: Calcium binding and translocation by the voltage-dependent anion channel: a possible regulatory mechanism in mitochondrial function.
abstract: Mitochondria play a central role in energy metabolism, Ca(2+) signalling, aging and cell death. To control cytosolic or mitochondrial Ca(2+) concentration, mitochondria possess several Ca(2+)-transport systems across the inner membrane. However, the pathway for Ca(2+) crossing the outer membrane has not been directly addressed. We report that purified voltage-dependent anion channel (VDAC) reconstituted into lipid bilayers or liposomes is highly permeable to Ca(2+). VDAC contains Ca(2+)-binding sites that bind Ruthenium Red (RuR), La(3+) and that RuR completely closed VDACs in single or multichannel experiments. Energized, freshly prepared mitochondria accumulate Ca(2+) (500-700 nmol/mg of protein), and subsequently released it. The release of Ca(2+) is accompanied by cyclosporin A-inhibited swelling, suggesting activation of permeability transition pore (PTP). RuR and ruthenium amine binuclear complex, when added to mitochondria after Ca(2+) accumulation has reached a maximal level and before PTP is activated, prevented the release of Ca(2+) and the accompanied mitochondrial swelling. RuR also prevented PTP opening promoted by atractyloside, an adenine nucleotide translocase inhibitor. These results suggest that VDAC, located in the mitochondrial outer membrane, controls Ca(2+) transport into and from the mitochondria, and that the inhibition of Ca(2+) uptake by RuR and La(3+) may result from their interaction with VDAC Ca(2+)-binding sites. Inhibition of PTP opening or assembly by RuR and ruthenium amine binuclear complex suggest the involvement of VDAC in PTP activity and/or regulation. The permeability of VDAC to Ca(2+) and its binding of Ca(2+), suggest that VDAC has a role in regulation of the mitochondrial Ca(2+) homoeostasis.
authors.affiliation: Department of Life Sciences and Zlotowski Center for Neuroscience, Ben Gurion University of the Negev, Beer Sheva, P.O.B. 653, 84105 Israel.
authors.forename: D
authors.initials: D
authors.lastname: Gincel
authors.forename: H
authors.initials: H
authors.lastname: Zaid
authors.forename: V
authors.initials: V
authors.lastname: Shoshan-Barmatz
mesh_terms: D000818:Animals; D000838:Anions; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D016572:Cyclosporine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D007473:Ion Channels / Q000378:metabolism*; D007700:Kinetics; D007811:Lanthanum / Q000494:pharmacology; D008051:Lipid Bilayers / Q000737:chemistry; D008081:Liposomes; D008928:Mitochondria / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism; D008954:Models, Biological; D015233:Models, Statistical; D018272:Porins / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D012430:Ruthenium Red / Q000494:pharmacology; D015398:Signal Transduction; D013997:Time Factors; D050994:Voltage-Dependent Anion Channels
chemical_list: D000838:Anions; D007473:Ion Channels; D008051:Lipid Bilayers; D008081:Liposomes; D018272:Porins; D050994:Voltage-Dependent Anion Channels; D012430:Ruthenium Red; D007811:Lanthanum; D016572:Cyclosporine; D002118:Calcium