Abstract
The role of the voltage-dependent anion channel (VDAC) in cell death was investigated using the expression of native and mutated murine VDAC1 in U-937 cells and VDAC inhibitors. Glutamate 72 in VDAC1, shown previously to bind dicyclohexylcarbodiimide (DCCD), which inhibits hexokinase isoform I (HK-I) binding to mitochondria, was mutated to glutamine. Binding of HK-I to mitochondria expressing E72Q-mVDAC1, as compared to native VDAC1, was decreased by ∼70% and rendered insensitive to DCCD. HK-I and ruthenium red (RuR) reduced the VDAC1 conductance but not that of E72Q-mVDAC1. Overexpression of native or E72Q-mVDAC1 in U-937 cells induced apoptotic cell death (80%). RuR or overexpression of HK-I prevented this apoptosis in cells expressing native but not E72Q-mVDAC1. Thus, a single amino-acid mutation in VDAC prevented HK-I- or RuR-mediated protection against apoptosis, suggesting the direct VDAC regulation of the mitochondria-mediated apoptotic pathway and that the protective effects of RuR and HK-I rely on their binding to VDAC.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- ANT:
-
adenine nucleotide translocase
- G-6-P:
-
glucose-6-phosphate
- Hepes:
-
N-(-hydroxyethyl])piperazine-N′-[2-ethanesulfonic acid]
- HK:
-
hexokinase
- HK-I:
-
hexokinase isoform I
- β-OG:
-
n-octyl-β-D-glucopyranoside
- PLB:
-
planar lipid bilayer
- RuR:
-
ruthenium red
- STS:
-
staurosporine
- VDAC:
-
voltage-dependent anion channel
- mVDAC:
-
murine VDAC
- rVDAC:
-
rat VDAC
References
Berridge MJ, Bootman MD and Lipp P (1998) Calcium – a life and death signal. Nature 395: 645–648
Kroemer G, Zamzami N and Susin SA (1997) Mitochondrial control of apoptosis. Immunol. Today 18: 44–51
Crompton M, Virji S, Doyle V, Johnson N and Ward JM (1999) The mitochondrial permeability transition pore. Biochem. Soc. Symp. 66: 167–179
Halestrap AP, Doran E, Gillespie JP and O'Toole A (2000) Mitochondria and cell death. Biochem. Soc. Trans. 28: 170–177
Vander Heiden MG, Chandel NS, Schumacker PT and Thompson CB (1999) Bcl-xL prevents cell death following growth factor withdrawal by facilitating mitochondrial ATP/ADP exchange. Mol. Cell 3: 159–167
Zoratti M and Szabo I (1995) The mitochondrial permeability transition. Biochim. Biophys. Acta 1241: 139–176
Gincel D, Zaid H and Shoshan-Barmatz V (2001) Calcium binding and translocation by the voltage-dependent anion channel: a possible regulatory mechanism in mitochondrial function. Biochem. J. 358: 147–155
Bernardi P (1999) Mitochondrial transport of cations: channels, exchangers, and permeability transition. Physiol. Rev. 79: 1127–1155
Cesura AM, Pinard E, Schubenel R, Goetschy V, Friedlein A, Langen H, Polcic P, Forte MA, Bernardi P and Kemp JA (2003) The voltage-dependent anion channel is the target for a new class of inhibitors of the mitochondrial permeability transition pore. J. Biol. Chem. 278: 49812–49818
Shoshan-Barmatz V and Gincel D (2003) The voltage-dependent anion channel: characterization, modulation, and role in mitochondrial function in cell life and death. Cell Biochem. Biophys. 39: 279–292
Brustovetsky N and Klingenberg M (1996) Mitochondrial ADP/ATP carrier can be reversibly converted into a large channel by Ca2+. Biochemistry 35: 8483–8488
Gincel D, Vardi N and Shoshan-Barmatz V (2002) Retinal voltage-dependent anion channel: characterization and cellular localization. Invest. Ophthalmol. Vis. Sci. 43: 2097–2104
Corbalan-Garcia S, Teruel JA and Gomez-Fernandez JC (1992) Characterization of ruthenium red-binding sites of the Ca2+-ATPase from sarcoplasmic reticulum and their interaction with Ca2+-binding sites. Biochem. J. 287: 767–774
Chen SR and MacLennan DH (1994) Identification of calmodulin, Ca2+ and ruthenium red-binding domains in the Ca2+ release channel (ryanodine receptor) of rabbit skeletal muscle sarcoplasmic reticulum. J. Biol. Chem. 269: 22698–22704
Charuk JH, Pirraglia CA and Reithmeier RA (1990) Interaction of ruthenium red with Ca2+-binding proteins. Anal. Biochem. 188: 123–131
Sasaki T, Naka M, Nakamura F and Tanaka T (1992) Ruthenium red inhibits the binding of calcium to calmodulin required for enzyme activation. J. Biol. Chem. 267: 21518–21523
Gunter TE, Buntinas L, Sparagna GC and Gunter KK (1998) The Ca2+ transport mechanisms of mitochondria and Ca2+ uptake from physiological-type Ca2+ transients. Biochim. Biophys. Acta 1366: 5–15
Deinum J, Wallin M, Kanje M and Lagercrantz C (1981) The effect of ruthenium red on the assembly and disassembly of microtubules and on rapid axonal transport. Biochim. Biophys. Acta 675: 209–213
Ying WL, Emerson J, Clarke MJ and Sanadi DR (1991) Inhibition of mitochondrial calcium ion transport by an oxo-bridged dinuclear ruthenium ammine complex. Biochemistry 30: 4949–4952
Bae JH, Park JW and Kwon TK (2003) Ruthenium red, inhibitor of mitochondrial Ca2+ uniporter, inhibits curcumin-induced apoptosis via the prevention of intracellular Ca2+ depletion and cytochrome c release. Biochem. Biophys. Res. Commun. 303: 1073–1079
Baek JH, Lee YS, Kang CM, Kim JA, Kwon KS, Son HC and Kim KW (1997) Intracellular Ca2+ release mediates ursolic acid-induced apoptosis in human leukemic HL-60 cells. Int. J. Cancer 73: 725–728
Ding WX, Shen HM and Ong CN (2001) Pivotal role of mitochondrial Ca2+ in microcystin-induced mitochondrial permeability transition in rat hepatocytes. Biochem. Biophys. Res. Commun. 285: 1155–1161
Bryson JM, Coy PE, Gottlob K, Hay N and Robey RB (2002) Increased hexokinase activity, of either ectopic or endogenous origin, protects renal epithelial cells against acute oxidant-induced cell death. J. Biol. Chem. 277: 11392–11400
Pastorino JG, Shulga N and Hoek JB (2002) Mitochondrial binding of hexokinase II inhibits Bax-induced cytochrome c release and apoptosis. J. Biol. Chem. 277: 7610–7618
Azoulay-Zohar H, Israelson A, Abu-Hamad S and Shoshan-Barmatz V (2004) In self-defence: hexokinase promotes voltage-dependent anion channel closure and prevents mitochondria-mediated apoptotic cell death. Biochem. J. 377: 347–355
Reymann S, Florke H, Heiden M, Jakob C, Stadtmuller U, Steinacker P, Lalk VE, Pardowitz I and Thinnes FP (1995) Further evidence for multitopological localization of mammalian porin (VDAC) in the plasmalemma forming part of a chloride channel complex affected in cystic fibrosis and encephalomyopathy. Biochem. Mol. Med. 54: 75–87
Song J and Colombini M (1996) Indications of a common folding pattern for VDAC channels from all sources. J. Bioenerg. Biomembr. 28: 153–161
Blachly-Dyson E, Song J, Wolfgang WJ, Colombini M and Forte M (1997) Multicopy suppressors of phenotypes resulting from the absence of yeast VDAC encode a VDAC-like protein. Mol. Cell. Biol. 17: 5727–5738
De Pinto V, al Jamal JA and Palmieri F (1993) Location of the dicyclohexylcarbodiimide-reactive glutamate residue in the bovine heart mitochondrial porin. J. Biol. Chem. 268: 12977–12982
Shafir I, Feng W and Shoshan-Barmatz V (1998) Dicyclohexylcarbodiimide interaction with the voltage-dependent anion channel from sarcoplasmic reticulum. Eur. J. Biochem. 253: 627–636
Shoshan-Barmatz V, Hadad N, Feng W, Shafir I, Orr I, Varsanyi M and Heilmeyer LM (1996) VDAC/porin is present in sarcoplasmic reticulum from skeletal muscle. FEBS Lett. 386: 205–210
Nakashima RA, Mangan PS, Colombini M and Pedersen PL (1986) Hexokinase receptor complex in hepatoma mitochondria: evidence from N, N′-dicyclohexylcarbodiimide-labeling studies for the involvement of the pore-forming protein VDAC. Biochemistry 25: 1015–1021
Godbole A, Varghese J, Sarin A and Mathew MK (2003) VDAC is a conserved element of death pathways in plant and animal systems. Biochim. Biophys. Acta 1642: 87–96
Tsujimoto Y and Shimizu S (2002) The voltage-dependent anion channel: an essential player in apoptosis. Biochimie 84: 187–193
Zalk R, Israelson A, Garty E, Azoulay-Zohar H and Shoshan-Barmatz V (2005) Oligomeric states of the voltage-dependent anion channel and cytochrome c release from mitochondria. Biochem. J. 386: 73–83
Shimizu S, Narita M and Tsujimoto Y (1999) Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 399: 483–487
Shimizu S, Konishi A, Kodama T and Tsujimoto Y (2000) BH4 domain of antiapoptotic Bcl-2 family members closes voltage-dependent anion channel and inhibits apoptotic mitochondrial changes and cell death. Proc. Natl. Acad. Sci. USA 97: 3100–3105
Vander Heiden MG, Li XX, Gottleib E, Hill RB, Thompson CB and Colombini M (2001) Bcl-xL promotes the open configuration of the voltage-dependent anion channel and metabolite passage through the outer mitochondrial membrane. J. Biol. Chem. 276: 19414–19419
Bota AD and Davies KJA (2001) Protein degradation in mitochondria: implications for oxidative stress, aging and disease: a novel etiological classification of mitochondrial proteolytic disorders. Mitochondrion 1: 33–49
Zheng Y, Shi Y, Tian C, Jiang C, Jin H, Chen J, Almasan A, Tang H and Chen Q (2004) Essential role of the voltage-dependent anion channel (VDAC) in mitochondrial permeability transition pore opening and cytochrome c release induced by arsenic trioxide. Oncogene 23: 1239–1247
Lee AC, Xu X, Blachly-Dyson E, Forte M and Colombini M (1998) The role of yeast VDAC genes on the permeability of the mitochondrial outer membrane. J. Membr. Biol. 161: 173–181
Bourgeron T, Chretien D, Rotig A, Munnich A and Rustin P (1992) Isolation and characterization of mitochondria from human B lymphoblastoid cell lines. Biochem. Biophys. Res. Commun. 186: 16–23
Gornall AG, Bardawill CJ and David MM (1949) Determination of serum proteins by means of the Biuret reaction. J. Biol. Chem. 177: 751–766
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Towbin H, Staehelin T and Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350–4354
McGahon AJ, Martin SJ, Bissonnette RP, Mahboubi A, Shi Y, Mogil RJ, Nishioka WK and Green DR (1995) The end of the (cell) line: methods for the study of apoptosis in vitro. Methods Cell Biol. 46: 153–185
Acknowledgements
This research was supported by a grant from the Israel Science Foundation, administrated by The Israel Academy of Science and Humanities. We gratefully acknowledge the assistance of Dr. Sara Sivan.
Author information
Authors and Affiliations
Corresponding author
Additional information
Edited by C Borner
Rights and permissions
About this article
Cite this article
Zaid, H., Abu-Hamad, S., Israelson, A. et al. The voltage-dependent anion channel-1 modulates apoptotic cell death. Cell Death Differ 12, 751–760 (2005). https://doi.org/10.1038/sj.cdd.4401599
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/sj.cdd.4401599
Keywords
This article is cited by
-
Comparative hemolymph proteomic analyses of the freezing and resistance-freezing Ostrinia furnacalis (Guenée)
Scientific Reports (2024)
-
Apoptotic proteins with non-apoptotic activity: expression and function in cancer
Apoptosis (2023)
-
Targeting the overexpressed mitochondrial protein VDAC1 in a mouse model of Alzheimer’s disease protects against mitochondrial dysfunction and mitigates brain pathology
Translational Neurodegeneration (2022)
-
Loss of primary cilia promotes mitochondria-dependent apoptosis in thyroid cancer
Scientific Reports (2021)
-
Structure, gating and interactions of the voltage-dependent anion channel
European Biophysics Journal (2021)
