Consumption of L-arginine mediated by Entamoeba histolytica L-arginase (EhArg) inhibits amoebicidal activity and nitric oxide production by activated macrophages
Keren Elnekave,
Rama Siman-Tov,
Serge Ankri,
Keren Elnekave
Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel
Search for more papers by this authorRama Siman-Tov
Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel
Search for more papers by this authorCorresponding Author
Serge Ankri
Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel
Serge Ankri, Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, P.O.B. 9649, 31096 Haifa, Israel (e-mail: [email protected]).Search for more papers by this authorKeren Elnekave,
Rama Siman-Tov,
Serge Ankri,
Keren Elnekave
Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel
Search for more papers by this authorRama Siman-Tov
Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel
Search for more papers by this authorCorresponding Author
Serge Ankri
Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel
Serge Ankri, Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, P.O.B. 9649, 31096 Haifa, Israel (e-mail: [email protected]).Search for more papers by this authorSUMMARY
In this study we discuss the cloning and expression of Entamoeba histolytica arginase (EhArg), an enzyme that catalyses the hydrolysis of l-arginine to l-ornithine and urea. l-norvaline, a competitive inhibitor of E. histolytica l-arginase, inhibits the growth of the parasite, which suggests that the catabolism of l-arginine mediated by EhArg is essential. Nitric oxide (NO) is an antimicrobial agent that inhibits some key enzymes in the metabolism of Entamoeba histolytica. NO is synthesized by activated macrophages from l-arginine, the substrate of NO synthase (NOS-II). We show that E. histolytica inhibits NO mediated amoebicidal activity of activated macrophages by consuming l-arginine present in the medium.
REFERENCES
- 1 Hamann L, Buss H & Tannich E. Tetracycline-controlled gene expression in Entamoeba histolytica. Mol Biochem Parasitol 1997; 84: 83–91.
- 2 Espinosa-Cantellano M & Martinez-Palomo A. Pathogenesis of intestinal amebiasis: from molecules to disease. Clin Microbiol Rev 2000; 13: 318–331.
- 3 Stanley SL Jr & Reed SL. Microbes and microbial toxins: paradigms for microbial–mucosal interactions. VI. Entamoeba histolytica: parasite–host interactions. Am J Physiol Gastrointest Liver Physiol 2001; 280: G1049–G1054.
- 4 Shiloh MU & Nathan CF. Reactive nitrogen intermediates and the pathogenesis of Salmonella and mycobacteria. Curr Opin Microbiol 2000; 3: 35–42.
- 5 Bogdan C, Rollinghoff M & Diefenbach A. The role of nitric oxide in innate immunity. Immunol Rev 2000; 173: 17–26.
- 6 Nathan C & Shiloh MU. Reactive oxygen and nitrogen intermediates in the relationship between mammalian hosts and microbial pathogens. Proc Natl Acad Sci USA 2000; 97: 8841–8848.
- 7 Lin JY & Chadee K. Macrophage cytotoxicity against Entamoeba histolytica trophozoites is mediated by nitric oxide from 1-arginine. J Immunol 1992; 148: 3999–4005.
- 8 Seydel KB, Smith SJ & Stanley SL Jr. Innate immunity to amebic liver abscess is dependent on gamma interferon and nitric oxide in a murine model of disease. Infect Immun 2000; 68: 400–402.
- 9 Siman-Tov R & Ankri S. Nitric oxide inhibits cysteine proteinases and alcohol dehydrogenase 2 of Entamoeba histolytica. Parasitol Res 2003; 89: 146–149.
- 10 Wang W, Keller K & Chadee K. Entamoeba histolytica modulates the nitric oxide synthase gene and nitric oxide production by macrophages for cytotoxicity against amoebae and tumour cells. Immunology 1994; 83: 601–610.
- 11 Kretschmer RR, Rico G & Gimenez JA. A novel anti-inflammatory oligopeptide produced by Entamoeba histolytica. Mol Biochem Parasitol 2001; 112: 201–209.
- 12 Mills CD. Macrophage arginine metabolism to ornithine/urea or nitric oxide/citrulline: a life or death issue. Crit Rev Immunol 2001; 21: 399–425.
- 13 Bansal V & Ochoa JB. Arginine availability, arginase, and the immune response. Curr Opin Clin Nutr Metab Care 2003; 6: 223–228.
- 14 Gobert AP, Daulouede S, Lepoivre M et al. L-Arginine availability modulates local nitric oxide production and parasite killing in experimental trypanosomiasis. Infect Immun 2000; 68: 4653–4657.
- 15 Eckmann L, Laurent F, Langford TD et al. Nitric oxide production by human intestinal epithelial cells and competition for arginine as potential determinants of host defense against the lumen-dwelling pathogen Giardia lamblia. J Immunol 2000; 164: 1478–1487.
- 16 Gobert AP, McGee DJ, Akhtar M et al. Helicobacter pylori arginase inhibits nitric oxide production by eukaryotic cells: a strategy for bacterial survival. Proc Natl Acad Sci USA 2001; 98: 13844–13849.
- 17 Diamond LS, Harlow DR & Cunnick CC. A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg 1978; 72: 431–432.
- 18 Narumi S, Finke JH & Hamilton TA. Interferon gamma and interleukin 2 synergize to induce selective monokine expression in murine peritoneal macrophages. J Biol Chem 1990; 265: 7036–7041.
- 19 Quintero MJ, Muro-Pastor AM, Herrero A & Flores E. Arginine catabolism in the cyanobacterium Synechocystis sp. Strain PCC 6803 involves the urea cycle and arginase pathway. J Bacteriol 2000; 182: 1008–1015.
- 20 Corraliza IM, Campo ML, Soler G & Modolell M. Determination of arginase activity in macrophages: a micromethod. J Immunol Meth 1994; 174: 231–235.
- 21 Kasprzak W & Majewska AC. Infectivity of Giardia sp. cysts in relation to eosin exclusion and excystation in vitro. Tropenmed Parasitol 1983; 34: 70–72.
- 22 Leippe M, Sievertsen HJ, Tannich E & Horstmann RD. Spontaneous release of cysteine proteinases but not of pore-forming peptides by viable Entamoeba histolytica. Parasitology 1995; 111: 569–574.
- 23 Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 1976; 72: 248–254.
- 24 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680–685.
- 25 Bellocco E, Di Salvo C, Lagana G et al. Purification and properties of ornithine carbamoyltransferase from loggerhead turtle liver. Physiol Res 2002; 51: 151–158.
- 26 Zuo X & Coombs GH. Amino acid consumption by the parasitic, amoeboid protists Entamoeba histolytica and E. invadens. FEMS Microbiol Lett 1995; 130: 253–258.
- 27 Jenkinson CP, Grody WW & Cederbaum SD. Comparative properties of arginases. Comp Biochem Physiol B Biochem Mol Biol 1996; 114: 107–132.
- 28 Molina MC, Stocker-Worgotter E, Turk R, Bajon C & Vicente C. Secreted, glycosylated arginase from Xanthoria parietina thallus induces loss of cytoplasmic material from Xanthoria photobionts. Cell Adhes Commun 1998; 6: 481–490.
- 29 Field J, Rosenthal B & Samuelson J. Early lateral transfer of genes encoding malic enzyme, acetyl-CoA synthetase and alcohol dehydrogenases from anaerobic prokaryotes to Entamoeba histolytica. Mol Microbiol 2000; 38: 446–455.
- 30 Nixon JE, Wang A, Field J et al. Evidence for lateral transfer of genes encoding ferredoxins, nitroreductases, NADH oxidase, and alcohol dehydrogenase 3 from anaerobic prokaryotes to Giardia lamblia and Entamoeba histolytica. Eukaryot Cell 2002; 1: 181–190.
- 31 Altschul SF, Gish W, Miller W, Myers EW & Lipman DJ. Basic local alignment search tool. J Mol Biol 1990; 215: 403–410.
- 32 Wilihoeft U, Campos-Gongora E, Touzni S et al. Introns of Entamoeba histolytica and Entamoeba dispar. Protist 2001; 152: 149–156.
- 33 Vockley JG, Goodman BK, Tabor DE et al. Loss of function mutations in conserved regions of the human arginase I gene. Biochem Mol Med 1996; 59: 44–51.
- 34 Hwang HJ, Kim EH & Cho YD. Isolation and properties of arginase from a shade plant, ginseng (Panax ginseng C.A. Meyer) roots. Phytochem 2001; 58: 1015–1024.
- 35 Kanda M, Ohgishi K, Hanawa T & Saito Y. Arginase of Bacillus brevis Nagano: purification, properties, and implication in gramicidin S biosynthesis. Arch Biochem Biophys 1997; 344: 37–42.
- 36 Dillon BJ, Holtsberg FW, Ensor CM, Bomalaski JS & Clark MA. Biochemical characterization of the arginine degrading enzymes arginase and arginine deiminase and their effect on nitric oxide production. Med Sci Monit 2002; 8: BR248–53.
- 37 Majumdar S, Gupta R & Dogra N. Interferon-gamma- and lipopolysaccharide-induced tumor necrosis factor-alpha is required for nitric oxide production: tumor necrosis factor-alpha and nitric oxide are independently involved in the killing of Mycobacterium microti in interferon-gamma- and lipopolysaccharide-treated J774A.1 cells. Folia Microbiol (Praha) 2000; 45: 457–463.
- 38 Chang CI, Liao JC & Kuo L. Macrophage arginase promotes tumor cell growth and suppresses nitric oxide-mediated tumor cytotoxicity. Cancer Res 2001; 61: 1100–1106.
- 39 Pierson DL, Cox SL & Gilbert BE. Human ornithine transcarbamylase. Purification and characterization of the enzyme from normal liver and the liver of a Reye's syndrome patient. J Biol Chem 1977; 252: 6464–6469.
- 40 Witthoft T, Eckmann L, Kim JM & Kagnoff MF. Enteroinvasive bacteria directly activate expression of iNOS and NO production in human colon epithelial cells. Am J Physiol 1998; 275: G564–G571.
- 41 Munder M, Eichmann K, Moran JM, Centeno F, Soler G & Modolell M. Th1/Th2-regulated expression of arginase isoforms in murine macrophages and dendritic cells. J Immunol 1999; 163: 3771–3777.
- 42 Knodler LA, Schofield PJ & Edwards MR. 1-arginine transport and metabolism in Giardia intestinalis support its position as a transition between the prokaryotic and eukaryotic kingdoms. Microbiology 1995; 141: 2063–2070.
- 43 Kashiwagi K, Miyamoto S, Suzuki F, Kobayashi H & Igarashi K. Excretion of putrescine by the putrescine–ornithine antiporter encoded by the potE gene of Escherichia coli. Proc Natl Acad Sci USA 1992; 89: 4529–4533.
- 44 Poolman B, Driessen AJ & Konings WN. Regulation of arginine–ornithine exchange and the arginine deiminase pathway in Streptococcus lactis. J Bacteriol 1987; 169: 5597–5604.
- 45 Selamnia M, Mayeur C, Robert V & Blachier F. Alpha-difluoromethylornithine (DFMO) as a potent arginase activity inhibitor in human colon carcinoma cells. Biochem Pharmacol 1998; 55: 1241–1245.
- 46 Eckmann L, Reed SL, Smith JR & Kagnoff MF. Entamoeba histolytica trophozoites induce an inflammatory cytokine response by cultured human cells through the paracrine action of cytolytically released interleukin-1 alpha. J Clin Invest 1995; 96: 1269–1279.
- 47 Seguin R, Mann BJ, Keller K & Chadee K. The tumor necrosis factor alpha-stimulating region of galactose-inhibitable lectin of Entamoeba histolytica activates gamma interferon-primed macrophages for amebicidal activity mediated by nitric oxide. Infect Immun 1997; 65: 2522–2527.
- 48 Mai UE, Perez-Perez GI, Wahl LM, Wahl SM, Blaser MJ & Smith PD. Soluble surface proteins from Helicobacter pylori activate monocytes/macrophages by lipopolysaccharide-independent mechanism. J Clin Invest 1991; 87: 894–900.
- 49 Roediger WE. Nitric oxide damage to colonocytes in colitis-by-association: remote transfer of nitric oxide to the colon. Digestion 2002; 65: 191–195.
- 50 Rozenfeld C, Martinez R, Figueiredo RT et al. Soluble factors released by Toxoplasma gondii-infected astrocytes down-modulate nitric oxide production by gamma interferon-activated microglia and prevent neuronal degeneration. Infect Immun 2003; 71: 2047–2057.
- 51 Janne J, Alhonen L, Hirvonen A & Wahlfors J. Molecular genetics of ornithine decarboxylase in human tumor cells. Adv Enzyme Regul 1989; 28: 81–91.
- 52 Thomas T & Thomas TJ. Polyamines in cell growth and cell death: molecular mechanisms and therapeutic applications. Cell Mol Life Sci 2001; 58: 244–258.
- 53 Arteaga-Nieto P, Lopez-Romero E, Teran-Figueroa Y et al. Entamoeba histolytica: purification and characterization of ornithine decarboxylase. Exp Parasitol 2002; 101: 215–222.
- 54 Chayen A, Mirelman D & Chayen R. Polyamines in Entamoeba invadens. Cell Biochem Funct 1984; 2: 115–118.
