Abstract
Ascariasis, an intestinal worm infection is caused by the parasite Ascaris lumbricoides and a report by World Health Organization (WHO) on soil transmitted helminths suggests that over one billion people are affected by Ascariasis. This disease is prevalent in developing countries, and in places of poor sanitation and unhygienic conditions. Even though anthelminthic drugs are available for the treatment of ascariasis, it is considered as a neglected tropical disease (NTD). Resistance of the parasite to the existing drugs necessitates a detailed study of its energy metabolism for identification of new drug targets. The catabolic pathway of the parasite is an evolved design well suited for parasitic life and obtains constant input from its host. Its energy metabolism is predominantly anaerobic. The parasite mitochondrion plays a key role as it lacks the functional tricarboxylic acid cycle (TCA cycle) and cytochrome oxidase activity. In adult ascarid mitochondrion, there is no external final electron acceptor and endogenously produced fumarate and 2-methyl branched-chain enoyl — CoAs function as the terminal electron acceptors instead of oxygen. In this study, elementary flux mode analysis (EFM), a metabolic pathway analysis tool has been applied to model energy metabolism of the parasite A. lumbricoides. This study identifies a set of enzymes that have been suggested to be essential for the survival of the parasite; the inhibition of these enzymes paralyzes the parasite. The key enzymes of glycolysis and the phosphoenolpyruvate carboxykinase-succinate pathway are identified as drug targets since the knock-out of any of these enzymes results in zero flux value for all EFM that have been identified.
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Crompton, D. W. (2001) Ascaris and ascariasis. Adv. Parasitol. 48: 285–375.
Harrow, I. D. and K. A. F. Gration (1985) Mode of action of the anthelmintics, morantel, pyrantel and levamisole on muscle cell membrane of the nematode Ascaris suum. Pestic. Sci. 16: 662–672.
Horton, J. (2000) Albendazole: A review of anthelmintic efficacy and safety in humans. Parasitol. 121: 113–132.
Kass, I. S., C. C. Wang, J. P. Walrond, and A. O. W. Stretton (1980) Avermectin B1a, a paralyzing anthelminthic that affects Interneurons and inhibitory motorneurons in Ascaris. Proc. Natl. Acad. Sci. USA 77: 6211–6215.
Boes, J. and A. B. Helwig (2000) Animal models of intestinal nematode infections of humans. Parasitol. 121: 97–111.
Anderson, T. J., M. E. Romero -Abal, and J. Jaenike (1993) Genetic structure and epidemiology of Ascaris populations: Patterns of host affiliation in Guatemala. Parasitol. 107: 319–334.
Zhu, X., R. B. Gasser, and N. B. Chilton (1998) Differences in the 5.8S rDNA sequences among ascarid nematodes. Int. J. Parasitol. 28: 617–622.
Wei, G. H., C. Y. Xu, H. Q. Song, S. J. Wei, M. J. Xu, R. Q. Lin, G. H. Zhao, S. Y. Huang, and X. Q. Zhu (2012) Comparitive analysis of the complete mitochondrial genomes of Ascaris lumbricoides and Ascaris Suum from human and pigs. Gene 492: 110–116.
Alba, J. E., M. N. Comia, G. Oyong, and F. Claveria (2009) Ascaris lumbricoides and Ascaris suum: A comparision of electrophoretic banding patterns of protein extracts from the reproductive organs and body wall. Vet. Archiv. 79: 281–291.
Bryant, C. (1994) Ancient biochemistries and evolution of parasites. Int. J. Parasitol. 24: 1089–1097.
Fairlamb, A. H. (1989) Novel biochemical pathways in parasitic protozoa. Parasitol. 99: 93–112.
Schomburg, I., A. Chang, C. Ebeling, M. Gremse, C. Heldt, G. Huhn, and D. Schomburg (2004) BRENDA, the enzyme database: Updates and major new developments. Nucleic Acids Res. 32: 431–433.
Caspi, R., T. Altman, J. M. Dale, K. Dreher, C. A. Fulcher, F. Gilham, P. Kaipa, A. S. Karthikeyan, A. Kothari, M. Krummenacker, M. Latendresse, L. A. Mueller, S. Paley, L. Popescu, A. Pujar, A. G. Shearer, P. Zhang, and P. D. Karp (2012) The Meta-Cyc database of metabolic pathways and enzymes and the Bio-Cyc collection of pathway/genome databases. Nucleic Acids Res. 40: 742–753.
Gasteiger, E., A. Gattiker, C. Hoogland, I. Ivanyi, R. D. Appel, and A. Bairoch (2003) EXPASY: the proteomics server for indepth protein knowledge and analysis. Nucleic Acids Res. 31: 3784–3788.
Barrett, J. (1994) Biochemistry of helminths. pp. 211–232. In: N. Chowdhury and I. Tada (eds.). Helminthology. Springer-Verlag, NY.
Komuniecki, R. and A. G. M. Tielens (2003) Carbohydrate and energy metabolism in parasitic helminths. pp. 339–358. In: J. J. Marr, T. W. Nilsen, and R. W. Komuniecki (eds.). Molecular Medical Parasitology. Academic Press, Amsterdam.
Van Vugt, F., P. Van Der Meer, and S. G. VanDen Berg (1979) The formation of propionate and acetate as terminal processes in the energy metabolism of the adult liver fluke Fasciola hepatica. Int. J. Biochem. 10: 11–18.
Suarez de Mata Z., M. E. Zarrant, R. Lizardo, and H. J. Saz (1983) 2-methylacetoacetyl-coenzyme A reductase from Ascaris muscle: Purification and properties. Arch. Biochem. Biophys. 226: 84–93.
Kanehisa, M. and S. Goto (2000) KEGG: Kyoto Encyclopedia of Genes and Genomes. Nucleic Acids Res. 28: 27–30.
Bueding, E. and H. J. Saz (1968) Pyruvate kinase and phosphoenolpyruvate carboxykinase activities of Ascaris muscle, Hymenolepis dimunita and Schistosoma mansoni. Comp. Biochem. Physiol. 20: 511–518.
Tielens, A. G. M. (1994) Energy generation in parasitic helminths. Parasitol. Today 10: 346–352.
Saz, H. J. and A. Weil (1961) Pathway of formation of α-Methylvalerate by Ascaris lumbricoides. J. Biol. Chem. 237: 2053–2056.
Saz, H. J. and A. Weil (1960) Pathway of formation of α-Methylbutyrate from carbohydrate by Ascaris lumbricoides muscle. J. Biol. Chem. 235: 914–918.
Palsson, B. O. (2006) Systems biology: Properties of reconstructed networks. Cambridge University press, Cambridge.
Schuster, S., T. Dandekar, and D. A. Fell (1999) Detection of elementary flux modes in biochemical networks: A promising tool for pathway analysis and metabolic engineering. Trends Biotechnol. 17: 53–60.
Schuster, S., D. A. Fell, and T. Dandekar (2000) A general definition of metabolic pathways useful for systematic organization and analysis of complex metabolic networks. Nat. Biotechnol. 18: 326–332.
Stelling, J., S. Klamt, K. Bettenbrock, S. Schuster, and E. D. Gilles (2002) Metabolic network structure determines key aspects of functionality and regulation. Nature 420: 190–193.
Cakir, T., C. S. Tacer, and K. O. Ulgen (2004) Metabolic pathway analysis of enzyme deficient human red blood cell. Biosyst. 78: 49–67.
Ozbayraktar, F. B. and K. O. Ulgen (2010) Drug target identification in sphingolipid metabolism by computational systems biology tools: Metabolic control analysis and metabolic pathway analysis. J. Biomed. Inform. 43: 537–549.
Klamt, S., J. Saez-Rodriguez, and E. D. Gilles (2007) Structural and functional analysis of Cellular networks with cellnetanalyser. BMC Sys. Biol. 1: 2.
Saz, H. J. and J. R. Arthur Vidrin (1959) The mechanism of formation of succinate and Propionate by Ascaris lumbricoides muscle. J. Biol. Chem. 234: 2001–2005.
Tielens, A. G. M., K. W. A. Van Grinsven, K. Henze, J. J. Van Hellemond, and W. Martin (2010) Acetate formation in the energy metabolism of parasitic helminths and protists. Int. J. parasitol. 40: 387–397.
Tielens, A. G. M., C. Rotte, J. J. Van Hellemond, and W. Martin (2002) Mitochondria as we don’t know them. Trends Biochem. Sci. 27: 564–572.
Omura, S., H. Miyadera, H. Ui, K. Shiomi, Y. Ymaguchi, R. Masuma, T. Nagamitsu, D. Takano, T. Sunazuka, A. Harder, H. Kolbl, M. Namikoshi, H. Miyoshi, K. Sakamoto, and K. Kita (2001) An anthelmintic compound nafuredin shows selective inhibition of complex I in helminth mitochondria. Proc. Natl. Acad. Sci. USA 98: 60–62.
Miyadera, H., K. Shiomi, H. Ui, Y. Yamaguchi, R. Masuma, H. Tomoda, H. Miyoshi, A. Osanai, K. Kita, and S. Omura (2002) Atpenins, potent and specific inhibitors of mitochondrial complex II (Succinate-Ubiquinone oxidoreductase). Proc. Natl. Acad. Sci. USA 100: 473–477.
Osanai, A., S. Harada, K. Sakamoto, H. Shimizu, D. K. Inaoka, and K. Kita (2009) Crystallization of mitochondrial rhodoquinol -fumarate reductase from the parasitic nematode Ascaris suum with the specific inhibitor flutolanil. Acta Crystalographica. 65: 941–944.
Ui, H., K. Shiomi, H. Suzuki, H. Hatano, H. Morimoto, Y. Yamaguchi, R. Masuma, T. Sunazuka, H. Shimamura, K. Sakamoto, K. Kita, H. Miyoshi, H. Tomoda, and S. Omura (2006) Verticiprone a new NADH-fumarate reductase inhibitor, produced by Verticillium sp. FK1 -1083. J. Antibiot. 59: 785–790.
Ui, H., K. Shiomi, H. Suzuki, H. Hatano, H. Morimoto, Y. Yamaguchi, R. Masuma, K. Sakamoto, K. Kita, H. Miyoshi, H. Tomoda, H. Tanaka, and S. Omura (2006) Paecilaminol, a new NADH-fumarate reductase inhibitor, produced by Paecilomyces sp. FKI-0550. J. Antibiot. 59: 591–596.
Bueding, E., H. J. Saz, and G. W. Farrow (1959) The effect of piperazine on succinate production by Ascaris lumbricoides. Br. J. Pharmacol. Chemother. 14: 497–500.
Del Castillo, J., W. C. Demello, and T. Morales Brit (1964) Mechanism of the paralyzing action of Piperazine on Ascaris lumbricoides muscle. Br. J. Pharmacol. Chemother. 22: 463–477.
Geary, T. G., C. A. Winterrowd, S. J. Alexander-Bowman, M. A. Favreau, S. C. Nulf, and R. D. Klein (1993) Ascaris suum: cloning of a cDNA encoding phosphoenolpyruvate carboxykinase. Exp. Parasitol. 77: 155–161.
Verlinde, C. L., V. Hannaert, C. Blonski, M. Willson, J. J. Perie, L. A. Fothergrill-Gilmore, F. R. Opperdoes, M. H. Gelb, W. G. Hol, and P. A. Michels (2001) Glycolysis as a target for the design of new anti-trypanosome drugs. Drug Resist. Updat. 4: 50–65.
Kulkarni, G., N. A. Sabris, and B. G. Harris (2004) Cloning, expression and purification of fumarase from the parasitic nematode Ascaris suum. Protein Expres. Purif. 33: 209–213.
Supowit, S. C. and B. G. Harris (1976) Ascaris suum hexokinase: purification and possible function in compartmentation of glucose 6-phosphate muscle. Biochim. Biophys. Acta 422: 48–59.
Staal, G. E., A. Kalff, E. C. Heesbeen, C. W. Van Veelen, and G. Rijksen (1987) Subunit composition, regulatory properties, and phosphorylation of phosphofructokinase from human gliomas. Cancer Res. 47: 5047–5051.
Harris, B. G., J. A. Starling, and H. W. Hofer (1982) Phosphofructokinase from Ascaris suum muscle. Methods Enzymol. 90: 44–49.
Starling, J. A., B. C. Allen, M. R. Kaeini, D. M. Payne, H. J. Blytt, H. W. Hofer, and B. G. Harris (1982) Phosphofructokinase from Ascaris suum. Purification and properties. J. Biol. Chem. 257: 3795–3800.
Kulkarni, G., G. S. Jagannatha Rao, N. G. Srinivasan, H. W. Hofer, P. M. Yuan, and B. G. Harris (1987) Ascaris suum phosphofructokinase: Phosphorylation by protein kinase and sequence of the phosphopepetide. J. Biol. Chem. 262: 32–34.
Rohrer, S. P., H. J. Saz, and T. Nowak (1986) purification and characterization of phosphoenolpyruvate carboxykinase from the parasitic helminth Ascaris suum. J. Biol. Chem. 261: 13049–13055.
Dharmarajan, L., C. L. Case, P. Dunten, and B. Mukhopadhyay (2008) Tyr235 of human cytosolic phosphoenolpyruvate carboxykinase influences catalysis through an anion-quadrupole interaction with phosphoenolpyruvate carboxylate. FEBS J. 275: 5810–5819.
Tejada, P., M. Sanchez -Moreno, M. Monteoliva, and H. Gomez- Banqueri (1987) Inhibition of Malate dehydrogenase enzymes by benzimidazole anthelminthics. Vet. Parasitol. 24: 269–274.
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Parvatham, K., Veerakumari, L. Drug target prediction using elementary mode analysis in Ascaris lumbricoides energy metabolism. Biotechnol Bioproc E 18, 491–500 (2013). https://doi.org/10.1007/s12257-012-0390-x
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DOI: https://doi.org/10.1007/s12257-012-0390-x