Characterization of VDAC1 as a plasma membrane NADH‐oxidoreductase

We have recently demonstrated that voltage dependent anion selective channel 1 (porin, isoform 1) can function as a transplasma membrane NADH:ferricyanide‐reductase. However, both the specific redox characteristics and the mechanism of electron transport in this enzyme presently remain unclear. Here we demonstrate that the redox capability of porin 1 is specific for ferricyanide as this same enzyme cannot reduce DCIP or cytochrome c in vitro. Furthermore, NADH‐dependent ferricyanide reduction associated with VDAC1 is not sensitive to the anion channel inhibitors DIDS and dextran sulfate. However, this activity can be inhibited by thiol chelators, suggesting that at least one of the two cysteine groups present in VDAC1 are critical for electron transfer. We propose a model on how electron transport may occur in VDAC1.

[1]  M. Baker,et al.  Apoptosis and redox homostasis: On a possible mechanism of action of Bcl-2 , 1998, Protoplasma.

[2]  V. De Pinto,et al.  VDAC1 Is a Transplasma Membrane NADH-Ferricyanide Reductase* , 2004, Journal of Biological Chemistry.

[3]  R. Casadio,et al.  New functions of an old protein: the eukaryotic porin or voltage dependent anion selective channel (VDAC). , 2003, The Italian journal of biochemistry.

[4]  J. Ly,et al.  Transplasma membrane electron transport: enzymes involved and biological function , 2003, Redox report : communications in free radical research.

[5]  F. L. Crane,et al.  Purification and Characterization of a Doxorubicin-inhibited NADH-quinone (NADH-ferricyanide) Reductase from Rat Liver Plasma Membranes* , 2002, The Journal of Biological Chemistry.

[6]  M. V. Heiden,et al.  Outer mitochondrial membrane permeability can regulate coupled respiration and cell survival. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[7]  D. Spray,et al.  Evidence for secretory pathway localization of a voltage-dependent anion channel isoform. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Baker,et al.  Plasma membrane NADH-oxidoreductase system: a critical review of the structural and functional data. , 2000, Antioxidants & redox signaling.

[9]  A. Messina,et al.  Porin Is Present in the Plasma Membrane Where It Is Concentrated in Caveolae and Caveolae-related Domains* , 1999, The Journal of Biological Chemistry.

[10]  J. van der Zee,et al.  Ascorbate Stimulates Ferricyanide Reduction in HL-60 Cells through a Mechanism Distinct from the NADH-dependent Plasma Membrane Reductase* , 1998, The Journal of Biological Chemistry.

[11]  E. Wolvetang,et al.  Effectors of the mammalian plasma membrane NADH-oxidoreductase system. Short-chain ubiquinone analogues as potent stimulators , 1996, Journal of bioenergetics and biomembranes.

[12]  M. Colombini,et al.  ATP Flux Is Controlled by a Voltage-gated Channel from the Mitochondrial Outer Membrane* , 1996, The Journal of Biological Chemistry.

[13]  E. Wolvetang,et al.  Apoptosis induced by inhibitors of the plasma membrane NADH-oxidase involves Bcl-2 and calcineurin. , 1996, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[14]  L. Henderson,et al.  NADPH oxidase of neutrophils. , 1996, Biochimica et biophysica acta.

[15]  F. Vaillant,et al.  Up-regulation of the plasma membrane oxidoreductase as a prerequisite for the viability of human Namalwa rho 0 cells. , 1994, The Journal of biological chemistry.

[16]  R. F. Cook,et al.  Characterization of caveolin-rich membrane domains isolated from an endothelial-rich source: implications for human disease , 1994, The Journal of cell biology.

[17]  N. Hilschmann,et al.  Channel active mammalian porin, purified from crude membrane fractions of human B lymphocytes or bovine skeletal muscle, reversibly binds the stilbene-disulfonate group of the chloride channel blocker DIDS. , 1994, Biological chemistry Hoppe-Seyler.

[18]  E. Wolvetang,et al.  The universality of bioenergetic disease: the role of mitochondrial mutation and the putative inter-relationship between mitochondria and plasma membrane NADH oxidoreductase. , 1994, Molecular aspects of medicine.

[19]  M. Sorgato,et al.  Channels in mitochondrial membranes: knowns, unknowns, and prospects for the future. , 1993, Critical reviews in biochemistry and molecular biology.

[20]  R. Benz,et al.  Characterization of SH groups in porin of bovine heart mitochondria. Porin cysteines are localized in the channel walls. , 1991, European journal of biochemistry.

[21]  R. A. Nakashima.  Hexokinase-binding properties of the mitochondrial VDAC protein: Inhibition by DCCD and location of putative DCCD-binding sites , 1989, Journal of bioenergetics and biomembranes.

[22]  W. Stühmer,et al.  Patch-clamping of the inner mitochondrial membrane reveals a voltage-dependent ion channel , 1987, Nature.

[23]  M. Colombini.  Regulation of the mitochondrial outer membrane channel, VDAC , 1987, Journal of bioenergetics and biomembranes.

[24]  P. Mangan,et al.  Ultrasteep voltage dependence in a membrane channel. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[25]  P. Pedersen,et al.  Hexokinase receptor complex in hepatoma mitochondria: evidence from N,N'-dicyclohexylcarbodiimide-labeling studies for the involvement of the pore-forming protein VDAC. , 1986, Biochemistry.

[26]  F. L. Crane,et al.  Transplasma-membrane redox systems in growth and development. , 1985, Biochimica et biophysica acta.

[27]  F. L. Crane,et al.  Bleomycin control of transplasma membrane redox activity and proton movement in HeLa cells. , 1985, Biochemical pharmacology.

[28]  P. Pedersen,et al.  Intracellular localization and properties of particulate hexokinase in the Novikoff ascites tumor. Evidence for an outer mitochondrial membrane location. , 1983, The Journal of biological chemistry.

[29]  K. Magleby,et al.  Single voltage-dependent chloride-selective channels of large conductance in cultured rat muscle. , 1983, Biophysical journal.

[30]  F. L. Crane,et al.  Adriamycin affects plasma membrane redox functions. , 1980, Biochemical and biophysical research communications.

[31]  M. Colombini.  A candidate for the permeability pathway of the outer mitochondrial membrane , 1979, Nature.