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Biological significance of phosphoenolpyruvate carboxykinase in a cestode parasite, Raillietina echinobothrida and effect of phytoestrogens on the enzyme from the parasite and its host, Gallus domesticus

Published online by Cambridge University Press:  09 May 2017

RAMNATH ,
ASIM KUMAR DUTTA ,
BARILIN DKHAR ,
VEENA TANDON  and
BIDYADHAR DAS
RAMNATH
Affiliation:
Department of Zoology, Biological Chemistry Laboratory, North-Eastern Hill University, Shillong-793022, Meghalaya, India
ASIM KUMAR DUTTA
Affiliation:
Department of Zoology, Biological Chemistry Laboratory, North-Eastern Hill University, Shillong-793022, Meghalaya, India
BARILIN DKHAR
Affiliation:
Department of Zoology, Biological Chemistry Laboratory, North-Eastern Hill University, Shillong-793022, Meghalaya, India
VEENA TANDON
Affiliation:
Biotech Park, Janki Puram Sector G, Kursi Road, Lucknow-226021, UP, India
BIDYADHAR DAS*
Affiliation:
Department of Zoology, Biological Chemistry Laboratory, North-Eastern Hill University, Shillong-793022, Meghalaya, India
*
*Corresponding author. Department of Zoology, Biological Chemistry Laboratory, North-Eastern Hill University, Shillong-793022, Meghalaya, India. E-mail: dasbidyadhar@gmail.com
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Summary

Phosphoenolpyruvate carboxykinase (PEPCK) is involved in glycolysis in the cestode parasite, Raillietina echinobothrida; whereas, it executes a gluconeogenic role in its host, Gallus domesticus. Because of its differing primary function in the cestode parasite and its host, this enzyme is regarded as a plausible anthelmintic target. Hence, the biological significance of PEPCK in the parasite was analysed using siRNA against PEPCK from R. echinobothrida (RePEPCK). In order to find out the functional differences between RePEPCK and GdPEPCK (PEPCK from its host, G. domesticus), PEPCK genes from both sources were cloned, over-expressed, characterized, and some properties of the purified enzymes were compared. RePEPCK and GdPEPCK showed a standard Michaelis-Menten kinetics with Kmapp of 46.9 and 22.9 µm, respectively, for phosphoenolpyruvate and Kmapp of 15.4 µm for oxaloacetate in GdPEPCK decarboxylation reaction. Here, we report antagonist behaviours of recombinant PEPCKs derived from the parasite and its host. In search of possible modulators for PEPCK, few phytoestrogens were examined on the purified enzymes and their inhibitory constants were determined and discussed. This study stresses the potential of these findings to validate PEPCK as the anthelmintic drug target for parasitism management.

Keywords

Gallus domesticusRaillietina echinobothridaPEPCKphytoestrogenssiRNA
Type
Research Article
Information
Parasitology , Volume 144 , Issue 9 , August 2017 , pp. 1264 - 1274
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Copyright
Copyright © Cambridge University Press 2017 

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References

REFERENCES

Aboobaker, A. A. and Blaxter, M. L. (2003). Use of RNA interference to investigate gene function in the human filarial nematode parasite Brugia malayi . Molecular and Biochemical Parasitology 129, 41–51.Google Scholar
Arinze, I. J. (1975). On the development of phosphoenolpyruvate carboxykinase and gluconeogenesis in guinea pig liver. Biochemical and Biophysical Research Communications 65, 184–189.CrossRefGoogle ScholarPubMed
Bai, Y., Zhang, Z., Jin, L., Kang, H., Zhu, Y., Zhang, L., Li, X., Ma, F., Zhao, L., Shi, B., Li, J., McManus, D. P., Zhang, W. and Wang, S. (2014). Genome-wide sequencing of small RNAs reveals a tissue-specific loss of conserved microRNA families in Echinococcus granulosus . BMC Genomics 15, 736.Google Scholar
Balan, M. D., Mcleod, M. J., Lotosky, W. R., Ghaly, M. and Holyoak, T. (2015). Inhibition and allosteric regulation of monomeric phosphoenolpyruvate carboxykinase by 3-Mercaptopicolinic acid. Biochemistry 54, 5878–5887.CrossRefGoogle ScholarPubMed
Ballard, F. J. and Hanson, R. W. (1967). Phosphoenolpyruvate carboxykinase and pyruvate carboxylase in developing rat liver. Biochemical Journal 104, 866–871.Google Scholar
Behm, C. A. and Bryant, C. (1975 a). Studies of regulatory metabolism in Moniezia expansa: the role of phosphoenolpyruvate carboxykinase. International Journal for Parasitology 5, 347–354.Google Scholar
Behm, C. A. and Bryant, C. (1975 b). Studies of regulatory metabolism in Moniezia expansa: general conditions. International Journal for Parasitology 5, 209–217.Google Scholar
Behm, C. A. and Bryant, C (1982). Phosphoenolpyruvate carboxykinase from Fasciola hepatica . International Journal for Parasitology 12, 271–278.Google Scholar
Bradford, M. M. (1976). A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry 72, 248–254.Google Scholar
Bryant, C. (1975). Carbon dioxide utilization, and the regulation of respiratory metabolic pathways in parasitic helminths. Advances in Parasitology 13, 35–69.CrossRefGoogle ScholarPubMed
Bryant, C. and Behm, A. C. (1989). Biochemical Adaptation in Parasites. Chapman and Hall: London, New York.Google Scholar
Choi, M. S., Jung, U. J., Yeo, J., Kim, M. J. and Lee, M. K. (2008). Genistein and daidzein prevent diabetes onset by elevating insulin level and altering hepatic gluconeogenic and lipogenic enzyme activities in non-obese diabetic (NOD) mice. Diabetes/Metabolism Research and Reviews 24, 74–81.Google Scholar
Colombo, G., Carlson, G. M. and Lardy, H. A. (1978). Phosphoenolpyruvate carboxykinase (guanosine tri-phosphate) from rat liver cytosol. Separation of homogeneous forms of the enzyme with high and low activity by chromatography on agarose-hexane-guanosine tri-phosphate. Biochemistry 17, 5321–5329.Google Scholar
Cornish, R. A., Wilkes, J. and Mettrick, D. F. (1981 a). A study of phosphoenolpyruvate carboxykinase from Moniliformis dubius (Acanthocephala). Molecular and Biochemical Parasitology 2, 151–166.Google Scholar
Cornish, R. A., Wilkes, J. and Mettrick, D. F. (1981 b). The levels of some metabolites in Moniliformis dubius (Acanthocephala). Journal of Parasitology 67, 754–756.Google Scholar
Das, B., Tandon, V. and Saha, N. (2004 a). Anthelmintic efficacy of Flemingia vestita (Fabaceae): alteration in the activities of some glycolytic enzymes in the cestode, Raillietina echinobothrida . Parasitology Research 93, 253–261.CrossRefGoogle ScholarPubMed
Das, B., Tandon, V. and Saha, N. (2004 b). Effects of phytochemicals of Flemingia vestita (Fabaceae) on glucose 6-phosphate dehydrogenase and enzymes of gluconeogenesis in a cestode (Raillietina echinobothrida). Comparative Biochemistry and Physiology 139C, 141–146.Google Scholar
Das, B., Tandon, V., Saxena, J. K., Joshi, S. and Singh, A. R. (2013). Purification and characterization of phosphoenolpyruvate carboxykinase from Raillietina echinobothrida, a cestode parasite of the domestic fowl. Parasitology 140, 136–146.Google Scholar
Das, B., Ramnath, , Dutta, A. K. and Tandon, V. (2015). Differential kinetics at PK/PEPCK branch point in the cestode, Raillietina echinobothrida . Experimental Parasitology 153, 151–159.Google Scholar
Dayer, M. R., Dayer, M. S. and Ghayour, O. (2013). Dynamic behavior of rat phosphoenolpyruvate carboxykinase inhibitors: new mechanism for enzyme inhibition. Protein Journal 32, 253–258.Google Scholar
DiTullio, N. W., Berkoff, C. E., Blank, B., Kostos, V., Stack, E. J. and Sounders, H. L. (1974). 3-mercaptopicolinic acid, an inhibitor of gluconeogenesis. Biochemical Journal 138, 387–394.Google Scholar
Dutta, A. K., Ramnath, , Tandon, V. and Das, B. (2016 a). Biocomputational analysis of phosphoenolpyruvate carboxykinase from Raillietina echinobothrida, a cestode parasite, and its interaction with possible modulators. Parasitology 143, 300–313.Google Scholar
Dutta, A. K., Ramnath, , Dkhar, B., Tandon, V. and Das, B. (2016 b). Cloning and expression of phosphoenolpyruvate carboxykinase from a cestode parasite and its solubilization from inclusion bodies using L-arginine. Protein Expression and Purification 125, 61–67.Google Scholar
Ford, L., Guiliano, D. B., Oksov, Y., Debnath, A. K., Liu, J., Williams, S. A., Blaxter, M. L. and Lustigman, S. (2005). Characterization of a novel filarial serine protease inhibitor, Ov-SPI-1, from Onchocerca volvulus, with potential multifunctional roles during development of the parasite. Journal of Biological Chemistry 280, 40845–40856.Google Scholar
Geldhof, P., Visser, A., Clark, D., Saunders, G., Britton, C., Gilleard, J., Berriman, M. and Knox, D. (2007). RNA interference in parasitic helminths: current situation, potential pitfalls and future prospects. Parasitology 134, 609–619.Google Scholar
Goodman, M. N. (1975). Effect of 3-mercaptopicolinic acid on gluconeogenesis and gluconeogenic metabolite concentrations in the isolated perfused rat liver. Biochemical Journal 150, 137–139.Google Scholar
Hebda, C. A. and Nowak, T. (1982 a). Phosphoenolpyruvate carboxykinase. Mn2+ and Mn2+ substrate complexes. Journal of Biological Chemistry 257, 5515–5522.CrossRefGoogle ScholarPubMed
Hebda, C. A. and Nowak, T. (1982 b). The purification, characterization, and activation of phosphoenolpyruvate carboxykinase from chicken liver mitochondria. Journal of Biological Chemistry 257, 5503–5514.CrossRefGoogle ScholarPubMed
Hoffmann, K. H., Mustafa, T. and Jorgensen, J. B. (1979). Role of pyruvate kinase, phosphoenolpyruvate carboxykinase, malic enzyme and lactate dehydrogenase in anaerobic energy metabolism of Tubifex spec. Journal of Comparative Physiology 130, 337–345.Google Scholar
Johnson, T. A. and Holyoak, T. (2012). The Ω-loop lid domain of phosphoenolpyruvate carboxykinase is essential for catalytic function. Biochemistry 51, 9547–9559.Google Scholar
Johnson, T. A., Mcleod, M. J. and Holyoak, T. (2016). Utilization of substrate intrinsic binding energy for conformational change and catalytic function in phosphoenolpyruvate carboxykinase. Biochemistry 55, 575–587.CrossRefGoogle ScholarPubMed
Katiyar, S. P., Jain, A., Dhanjal, J. K. and Sundar, D. (2015). Mixed inhibition of cPEPCK by genistein, using an extended binding site located adjacent to its catalytic cleft. PloS ONE 10, e0141987.Google Scholar
Lee, M. H., Hebda, C. A. and Nowak, T. (1981). The role of cations in avian liver phosphoenolpyruvate carboxykinase catalysis. Activation and regulation. Journal of Biological Chemistry 256, 12793–12801.Google Scholar
Lustigman, S., Zhang, J., Liu, J., Oksov, Y. and Hashmi, S. (2004). RNA interference targeting cathepsin L and Z-like cysteine proteases of Onchocerca volvulus confirmed their essential function during L3 molting. Molecular and Biochemical Parasitology 138, 165–170.CrossRefGoogle ScholarPubMed
MacDonald, M. J. (1979). Evidence that Fe2+ complexes of 3-aminopicolinate and 3-mercaptopicolonate activate and inhibit phosphoenolpyruvate carboxykinase. Biochemical and Biophysical Research Communications 90, 741–749.Google Scholar
Moon, T. W., Mustafa, T., Hulbert, W. C., Podesta, R. B. and Metrrik, D. F. (1977). The phosphoenolpyruvate branch point in adult Hymenolepis diminuta (Cestoda): a study of pyruvate kinase and phosphoenolpyruvate carboxykinase. Journal of Experimental Zoology 200, 325–326.Google Scholar
Naguleswaran, A., Spicher, M., Vonlaufen, N., Ortega-Mora, L. M., Torgerson, P., Gottstein, B. and Hempill, A. (2006). In vitro metacestodicidal activities of genistein and other isoflavones against Echinococcus multilocularis and Echinococcus granulosus . Antimicrobial Agents and Chemotherapy 50, 3770–3778.Google Scholar
Nelson, D. L. and Cox, M. M. (2012). Lehninger Principles of Biochemistry, 6th Edn. WH Freeman and Company, USA.Google Scholar
Noce, P. S. and Utter, M. F. (1975). Decarboxylation of oxalacetate to pyruvate by purified avian liver phosphoenolpyruvate carboxykinase. Journal of Biological Chemistry 250, 9099–9105.Google Scholar
Nordlie, R. C. and Lardy, H. A. (1963). Mammalian liver phospoenolpyruvate carboxykinase activities. Journal of Biological Chemistry 238, 2259–2263.Google Scholar
Nsengimana, J., Bauters, L., Haegeman, A. and Gheysen, G. (2013). Silencing of Mg-pat-10 and Mg-unc-87 in the plant parasitic nematode Meloidogyne graminicola using siRNAs. Agriculture 3, 567–578.Google Scholar
Pfarr, K., Heider, U. and Hoerauf, A. (2006). RNAi mediated silencing of actin expression in adult Litomosoides sigmodontis is specific, persistent and results in a phenotype. International Journal for Parasitology 36, 661–669.Google Scholar
Pierson, L., Mousley, A., Devine, L., Marks, N. J., Day, T. A. and Maule, A. G. (2010). RNA interference in a cestode reveals specific silencing of selected highly expressed gene transcripts. International Journal for Parasitology 40, 605–615.Google Scholar
Prichard, R. K. (1976). Regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase activity in adult Fasciola hepatica (Trematoda). International Journal of Parasitology 6, 227–233.Google Scholar
Prichard, R. K. and Schofield, P. J. (1968). The metabolism of phosphoenolpyruvate and pyruvate in the adult fluke Fasciola hepatica . Biochimica et Biophysica Acta 170, 63–76.CrossRefGoogle Scholar
Reynolds, C. H. (1980). Phosphoenolpyruvate carboxykinase from the rat and from the tapeworm Hymenolepis diminuta. Effects of inhibitors and transition-metals on the carboxylation reaction. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 65, 481–487.Google Scholar
Roberts, L. S. (1983). Carbohydrate metabolism. In Biology of the Eucestoda, Vol II (ed. Arme, C. and Pappas, P. W.), pp. 343–390. Academic Press, New York.Google Scholar
Rognstad, R. (1981). Manganese effects on gluconeogenesis. Journal of Biological Chemistry 256, 1608–1610.Google Scholar
Rohrer, S. P., Saz, H. J. and Nowak, T. (1986). Purification and characterization of phosphoenolpyruvate carboxykinase from the parasitic helminth Ascaris suum . Journal of Biological Chemistry 261, 13049–13055.Google Scholar
Saz, H. J. and Lescure, O. L. (1969). The functions of phosphoenolpyruvate carboxykinase and malic enzyme in the anaerobic formation of succinate by Ascaris lumbricoides . Comparative Biochemistry and Physiology 30, 49–60.Google Scholar
Schramm, V. L., Fullin, F. A. and Zimmerman, M. D. (1981). Kinetic studies of the interaction of substrates, Mn2+, and Mg2+ with the Mn2+-sensitive and -insensitive forms of phosphoenolpyruvate carboxykinase. Journal of Biological Chemistry 256, 10803–10808.Google Scholar
Seenappa, V., Das, B., Joshi, M. B. and Satyamoorthy, K. (2016). Context dependent regulation of human phosphoenolpyruvate carboxykinase isoforms by DNA promoter methylation and RNA stability. Journal of Cellular Biochemistry 117, 2506–2520.CrossRefGoogle ScholarPubMed
Singh, P. K., Kushwaha, S., Mohd, S., Pathak, M. and Misra-Bhattacharya, S. (2013). In vitro gene silencing of independent phosphoglycerate mutase (iPGM) in the filarial parasite Brugia malayi . Infectious Diseases of Poverty 2, 5.Google Scholar
Smyth, J. D. and McManus, D. P. (1989). The Physiology and Biochemistry of Cestodes. Cambridge University Press, Cambridge, New York, Melbourne.Google Scholar
Soulsby, E. J. L. (1982). Helminths, Arthropods and Protozoa of Domesticated Animals, 7th Edn. ELBS and Bailliere Tindall, London, UK.Google Scholar
Tandon, V. and Das, B. (2007). In vitro testing of anthelmintic efficacy of Flemingia vestita (Fabaceae) on carbohydrate metabolism in Raillietina echinobothrida . Methods 42, 330–338.Google Scholar
Tandon, V., Pal, P., Roy, B., Rao, H. S. P. and Reddy, K. S. (1997). In vitro anthelmintic activity of root tuber extract of Flemingia vestita, an indigenous plant in Shillong, India. Parasitology Research 83, 492–498.Google Scholar
Utter, M. F. and Kurahashi, K. (1953). Mechanism of action of oxaloacetate carboxylase from liver. Journal of the American Chemical Society 75, 785–787.Google Scholar
Wilkes, J., Cornish, R. A. and Mettrick, D. F. (1981). Purification and properties of phosphoenolpyruvate carboxykinase from Hymenolepis diminuta (Cestoda). Journal of Parasitology 67, 832–840.Google Scholar
Wilkes, J., Cornish, R. A. and Mettrick, D. F. (1982). Purification and properties of phosphoenolpyruvate carboxykinase from Ascaris suum . International Journal for Parasitology 12, 163–171.Google Scholar
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