Effects of heavy metal cations on the mitochondrial ornithine/citrulline transporter reconstituted in liposomes

The effect of heavy metal cations on the mitochondrial ornithine/citrulline transporter was tested in proteoliposomes reconstituted with the protein purified from rat liver. The transport activity was measured as [3H]ornithine uptake in proteoliposomes containing internal ornithine (ornithine/ornithine antiport mode) or as [3H]ornithine efflux in the absence of external substrate (ornithine/H+ transport mode). 0.1 mM Cu2+, Pb2+, Hg2+, Cd2+ and Zn2+ strongly inhibited (more than 85%) the antiport; whereas Mn2+, Co2+ and Ni2+ inhibited less efficiently (25, 47 and 69%, respectively). The IC50 values of the transporter for the different metal ions ranged from 0.71 to 350 μM. Co2+ and Ni2+ also inhibited the [3H]ornithine efflux whereas Cu2+, Pb2+, Hg2+, Cd2+ and Zn2+ stimulated the [3H]ornithine efflux. The stimulation of the [3H]ornithine efflux by Cu2+ and Cd2+ (as well as by Pb2+, Hg2+ and Zn2+) was not prevented by NEM and was reversed by DTE. These features indicated that the inhibition of the antiport was due to the interaction of the Cu2+, Pb2+, Hg2+, Cd2+ and Zn2+ with a population of SH groups, of the transporter, responsible for the inhibition of the physiological function; whereas the stimulation of [3H]ornithine efflux was due to the induction of a pore-like function of the transporter caused by interaction of cations with a different population of SH groups. Differently, the inhibition of the ornithine transporter by Ni2+, Co2+ or Mn2+ was caused by interaction with the substrate binding site, as indicated by the competitive or mixed inhibition.

[1]  M. Norenberg,et al.  Signaling factors in the mechanism of ammonia neurotoxicity , 2009, Metabolic Brain Disease.

[2]  Edward I. Solomon,et al.  Structural and Functional Aspects of Metal Sites in Biology , 1997 .

[3]  F. Palmieri,et al.  Relationships of Cysteine and Lysine residues with the substrate binding site of the mitochondrial ornithine/citrulline carrier: an inhibition kinetic approach combined with the analysis of the homology structural model. , 2005, Biochimica et biophysica acta.

[4]  F. Thévenod.  Catch me if you can! Novel aspects of cadmium transport in mammalian cells , 2010, BioMetals.

[5]  E. Stadtman,et al.  Mitochondria play no roles in Mn(II)-induced apoptosis in HeLa cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[6]  S. Almashanu,et al.  Hyperornithinaemia-hyperammonaemia-homocitrullinuria syndrome is caused by mutations in a gene encoding a mitochondrial ornithine transporter , 1999, Nature Genetics.

[7]  A. Mathee,et al.  Levels of toxic and essential metals in maternal and umbilical cord blood from selected areas of South Africa--results of a pilot study. , 2009, Journal of environmental monitoring : JEM.

[8]  H. Bellamy,et al.  Crystallographic location of two Zn(2+)-binding sites in the avian cytochrome bc(1) complex. , 2000, Biochimica et biophysica acta.

[9]  T. Dierks,et al.  Pore-like and carrier-like properties of the mitochondrial aspartate/glutamate carrier after modification by SH-reagents: evidence for a performed channel as a structural requirement of carrier-mediated transport. , 1990, Biochimica et biophysica acta.

[10]  C. Heberger,et al.  Functional reconstitution of carrier proteins by removal of detergent with a hydrophobic ion exchange column. , 1986, Biochimica et biophysica acta.

[11]  N. Pavón,et al.  Copper induces permeability transition through its interaction with the adenine nucleotide translocase , 2007, Cell biology international.

[12]  Ferdinando Palmieri,et al.  The mitochondrial transporter family (SLC25): physiological and pathological implications , 2004, Pflügers Archiv.

[13]  Z. Shaikh,et al.  Lead nephrotoxicity and associated disorders: biochemical mechanisms. , 1992, Toxicology.

[14]  A. Konstantinov,et al.  Zinc ions as cytochrome c oxidase inhibitors: two sites of action , 2005, Biochemistry (Moscow).

[15]  Lorenzo Galluzzi,et al.  Mitochondrial membrane permeabilization in cell death. , 2007, Physiological reviews.

[16]  M. Zoratti,et al.  Mitochondrial permeability transitions: how many doors to the house? , 2005, Biochimica et biophysica acta.

[17]  R. Krämer,et al.  Functional properties of the reconstituted phosphate carrier from bovine heart mitochondria: evidence for asymmetric orientation and characterization of three different transport modes. , 1993, Biochimica et biophysica acta.

[18]  P. Bernardi,et al.  The mitochondrial permeability transition from yeast to mammals , 2010, FEBS letters.

[19]  Ferdinando Palmieri,et al.  Diseases caused by defects of mitochondrial carriers: a review. , 2008, Biochimica et biophysica acta.

[20]  V. Iacobazzi,et al.  Mitochondrial metabolite carrier proteins: purification, reconstitution, and transport studies. , 1995, Methods in enzymology.

[21]  J. Pourahmad,et al.  A comparison of hepatocyte cytotoxic mechanisms for Cu2+ and Cd2+. , 2000, Toxicology.

[22]  Pam Tucker,et al.  Toxicological Profile for Cadmium , 2012 .

[23]  F. Palmieri,et al.  The purified and reconstituted ornithine/citrulline carrier from rat liver mitochondria catalyses a second transport mode: ornithine+/H+ exchange. , 1999 .

[24]  Guibin Jiang,et al.  Biomonitoring: an appealing tool for assessment of metal pollution in the aquatic ecosystem. , 2008, Analytica chimica acta.

[25]  J. Rooney.  The role of thiols, dithiols, nutritional factors and interacting ligands in the toxicology of mercury. , 2007, Toxicology.

[26]  F. Palmieri,et al.  The purified and reconstituted ornithine/citrulline carrier from rat liver mitochondria: electrical nature and coupling of the exchange reaction with H+ translocation. , 1997, The Biochemical journal.

[27]  S. Chandna,et al.  Putative partners in Bax mediated cytochrome-c release: ANT, CypD, VDAC or none of them? , 2009, Mitochondrion.

[28]  Xianglin Shi,et al.  Metal-induced toxicity, carcinogenesis, mechanisms and cellular responses , 2004, Molecular and Cellular Biochemistry.

[29]  A. Driessen,et al.  What we can learn from the effects of thiol reagents on transport proteins. , 1992, Biochimica et biophysica acta.

[30]  Linjiang Li,et al.  Cadmium directly induced the opening of membrane permeability pore of mitochondria which possibly involved in cadmium-triggered apoptosis. , 2003, Toxicology.

[31]  T. Dierks,et al.  The mitochondrial carnitine carrier: characterization of SH-groups relevant for its transport function. , 1992, Biochimica et biophysica acta.

[32]  Lihua He,et al.  Lead and Calcium Produce Rod Photoreceptor Cell Apoptosis by Opening the Mitochondrial Permeability Transition Pore* , 2000, The Journal of Biological Chemistry.

[33]  A. Massadeh,et al.  Simultaneous Determination of Cd, Pb, Cu, Zn, and Se in Human Blood of Jordanian Smokers by ICP-OES , 2009, Biological Trace Element Research.

[34]  Seon-Hee Oh,et al.  A rapid and transient ROS generation by cadmium triggers apoptosis via caspase-dependent pathway in HepG2 cells and this is inhibited through N-acetylcysteine-mediated catalase upregulation. , 2006, Toxicology and applied pharmacology.

[35]  M. Spielmann,et al.  Cd2+-induced swelling-contraction dynamics in isolated kidney cortex mitochondria: role of Ca2+ uniporter, K+ cycling, and protonmotive force. , 2005, American journal of physiology. Cell physiology.

[36]  T. Dierks,et al.  The mitochondrial aspartate/glutamate and ADP/ATP carrier switch from obligate counterexchange to unidirectional transport after modification by SH-reagents. , 1990, Biochimica et biophysica acta.

[37]  Rudolfs K. Zalups,et al.  Molecular and ionic mimicry and the transport of toxic metals. , 2005, Toxicology and applied pharmacology.

[38]  John E. Walker,et al.  The Mitochondrial Ornithine Transporter , 2003, Journal of Biological Chemistry.

[39]  C. Indiveri,et al.  Chemical modification of the mitochondrial ornithine/citrulline carrier by SH reagents: effects on the transport activity and transition from carrier to pore-like function. , 2003, Biochimica et biophysica acta.

[40]  L. Thorpe,et al.  A Biomonitoring Study of Lead, Cadmium, and Mercury in the Blood of New York City Adults , 2007, Environmental health perspectives.

[41]  F. Palmieri,et al.  Identification and purification of the ornithine/citrulline carrier from rat liver mitochondria. , 1992, European journal of biochemistry.

[42]  S. Morris,et al.  Hormonal induction of hepatic mitochondrial ornithine/citrulline transporter mRNA. , 2002, Biochemical and biophysical research communications.

[43]  A. Albores,et al.  Cadmium and mercury toxicity in a human fetal hepatic cell line (WRL-68 cells). , 1995, Toxicology.

[44]  R. Unwin,et al.  Heavy metal poisoning: the effects of cadmium on the kidney , 2010, BioMetals.

[45]  R. Prasad,et al.  Evaluation of copper toxicity in isolated human peripheral blood mononuclear cells and it's attenuation by zinc: ex vivo , 2006, Molecular and Cellular Biochemistry.

[46]  Bing Zhou,et al.  Copper and manganese induce yeast apoptosis via different pathways. , 2007, Molecular biology of the cell.

[47]  P. Grieve,et al.  A simple technique for eliminating interference by detergents in the Lowry method of protein determination. , 1975, Analytical biochemistry.

[48]  J. Kaplan,et al.  Copper Transport in Mammalian Cells: Special Care for a Metal with Special Needs* , 2009, The Journal of Biological Chemistry.

[49]  A. Lavoinne,et al.  Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycle. , 2003, European journal of biochemistry.

[50]  Rudolfs K. Zalups,et al.  Molecular interactions with mercury in the kidney. , 2000, Pharmacological reviews.

[51]  O. Steward,et al.  Zn2+ Induces Permeability Transition Pore Opening and Release of Pro-apoptotic Peptides from Neuronal Mitochondria* , 2001, The Journal of Biological Chemistry.

[52]  A. Rosenzweig,et al.  Crystal structure of yeast Sco1 , 2006, JBIC Journal of Biological Inorganic Chemistry.