Advertisement

Journal of Parasitic Diseases

, Volume 33, Issue 1–2, pp 57–64 | Cite as

Characterization of the glutamate dehydrogenase activity of Gigantocotyle explanatum and Gastrothylax crumenifer (Trematoda: Digenea)

Original Article

Abstract

Glutamate dehydrogenase (GLDH) (EC 1.4.1.3) is a ubiquitous enzyme, which is present at the protein and carbohydrate metabolism crossroads. The enzyme activity was investigated in biliary and rumen amphistomes, Gigantocotyle explanatum and Gastrothylax crumenifer, respectively, infecting the Indian water buffalo Bubalus bubalis. The enzyme activity was consistently higher in G. explanatum as compared to G. crumenifer, where NAD(H) was utilized as coenzyme and the pH optima was recorded at 8. The Km and Vmax values for α-ketoglutarate were 2.1 mM and 9.09 units in G. explanatum, whereas 3.03 mM and 1.90 units in G. crumenifer, respectively. Among the allosteric modulator nucleotides, AMP, ADP, ATP, GMP, CMP and UMP, only AMP enhanced GLDH activity in G. crumenifer while ADP was stimulatory in G. explanatum. The amino acid leucine stimulated the GLDH activity in both the amphistomes while alanine was stimulatory only in G. crumenifer. Pronounced interspecific differences in response to different metabolic inhibitors like diethyldithiocarbamate, semicarbazide hydrochloride and mercurial ions were also observed. The osmotic stress alters the enzyme activity, particularly in hypertonic saline the GLDH activity increased significantly (p < 0.01) in G. explanatum, while insignificant effects were observed in rumen dwelling G. crumenifer. Histoenzymology revealed region/tissue specific distribution of GLDH with prominent staining in tissues like vitellaria, lymph system and tegument/subtegument, thus showing specific distribution of GLDH indicating differential metabolic state. Such intergeneric differences in GLDH activity could also be a consequence of occupying different microenvironments within the same host.

Keywords

Glutamate dehydrogenase Kinetics Allosteric modulators Osmotic stress Gigantocotyle explanatum Gastrothylax crumenifer 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Barrett J (1981) Biochemistry of Parasitic Helminths. Macmillan Publishers Ltd., LondonGoogle Scholar
  2. Cutillas C, Rodriguez B, Guevara DC (1992) Isoenzymatic pattern and structure of glutamate dehydrogenase from Ascaris suum. J Helminthol, 66:310–312CrossRefPubMedGoogle Scholar
  3. Czerkawski JW (1986) An introduction to rumen studies. Pergamon Press. OxfordGoogle Scholar
  4. Dominguez ND, Rodriguez-Acosta A (1996) Glutamate dehydrogenase antigen detection in Plasmodium falciparum infections. Korean J Parasitol, 34(4):239–246CrossRefPubMedGoogle Scholar
  5. Dunn TS, Nizami WA, Hanna REB (1985) Studies on the ultrastructure and histochemistry of the lymph system in three species of amphistome (Trematoda: Digenea) Gigantocotyle explanatum, Gastrothylax crumenifer and Srivastava indica from the Indian water buffalo Bubalus bubalis. J Helminthol, 59:1–18CrossRefPubMedGoogle Scholar
  6. Florkin M, Schoffeniels E (1969) Molecular Approach to Ecology. Academic Press, New York and LondonGoogle Scholar
  7. Frieden C (1959) Glutamate dehydrogenase II. The effect of various nucleotides on the association dissociation and kinetic properties. J Biol Chem, 234:815–820PubMedGoogle Scholar
  8. Krauth-Siegel RL, Müller JG, Lottspeich F, Schirmer RH (1996) Glutathione reductase and glutamate dehydrogenase of Plasmodium falciparum, the causative agent of tropical malaria. Eur J Biochem, 235:345–350CrossRefPubMedGoogle Scholar
  9. Krvavica S, Thomen H, Prosenjak M, Kucan D (1967) The presence and activity of glutamate dehydrogenase, glutamine synthetase and transaminases in Fasciola hepatica, Dicrocoelium lanceolatum and Paramphistomum cervi. Second International liver fluke Colloquim. Wageningen (Publ.), The NetherlandsGoogle Scholar
  10. Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc, 56:658–666CrossRefGoogle Scholar
  11. Lojda Z, Gossrau R, Schiebler TH (1979) Enzyme histochemistry - A laboratory manual. Springer-Verlag, Berlin, Heidelberg, New YorkGoogle Scholar
  12. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  13. McGivan JD, Chappell JB (1973) On the metabolic function of glutamate dehydrogenase in rat liver. FEBS letters 52:1–7CrossRefGoogle Scholar
  14. McNeil KM, Hutchinson WF (1971) The tricarboxylic acid cycle enzymes in the adult dog heart worm Dirofilaria immitis. Comp Biochem Physiol, 38:493–500CrossRefGoogle Scholar
  15. Mustafa T, Komuniecki R, Mettrick DF (1978) Cytosolic glutamate dehydrogenase in adult Hymenolepis diminuta (cestoda). Comp Biochem Physiol, 61B:219–222Google Scholar
  16. Olson JA, Anfinson CB (1953) Kinetic and equilibrium studies on crystalline L-glutamic acid dehydrogenase. J Biol Chem, 202:841–856PubMedGoogle Scholar
  17. Phiri IK, Phiri AM, Harrison LJ (2007) The serum glucose and beta-hydroxybutyrate levels in sheep with experimental Fasciola hepatica and Fasciola gigantica infection. Vet Parasitol, 143:287–293CrossRefPubMedGoogle Scholar
  18. Mohan Rao NK, Husain MM (1979) L-glutamate dehydrogenase activity of axenic Hartmanella culbertsoni. Ind J Parasitol, 3:5–23.Google Scholar
  19. Schmidt E (1974) Glutamate dehydrogenase UV assay. In: Methods of Enzymatic Analysis. Bergmeyer HU (Ed.), Academic Press, New York and London, pp. 650–656Google Scholar
  20. Schuster R, Dell K, Nockler K, Voster J, Schwartz-Porsche D, Haider W (2003) Liver enzyme activity and histological changes in the liver of silver foxes (Vulpes vulpes fulva) experimentally infected with opisthorchiid liver flukes. A contribution to the pathogenesis of opisthorchiidosis. Parasitol Res, 89(5):414–418PubMedGoogle Scholar
  21. Siddiqi AH, Islam MW, Nizami WA (1975) Osmotic and ionic behaviour of some digenetic trematodes. Comp Biochem Physiol, 51:929–935CrossRefGoogle Scholar
  22. Skuce PJ, Stewart EM, Smith WD, Knox DP (1999) Cloning and characterization of glutamate dehydrogenase (GDH) from the gut of Haemonchus contortus. Parasitology 118:297–304CrossRefPubMedGoogle Scholar
  23. Smith EL, Austin BM, Bluementhal KM, Nyc JF (1975) Glutamate dehydrogenases. In: The Enzymes. Boyer PD (Ed.), Vol. II, Academic Press, New York, pp. 294–367Google Scholar
  24. Sokal RR, Rohlf FJ (1981) Biometry: The principles and practice of statistics in biological research. WH Freeman and Company, New YorkGoogle Scholar
  25. Stephen HB, Alan K, Lawrence LD, David H, Richard JH (1978) NADP-specific glutamate dehydrogenase in Metridium senile (L.). Comp Biochem Physiol, 61:185–187CrossRefGoogle Scholar
  26. Vilas R, Paniagua E, Sanmartin ML (2002) Allozyme markers for the identification of Lecithochirium rufoviridae (Trematoda: Hemiuridae), parasites of Conger conger and Auguilla anguilla from Atlantic Spanish waters. J Parasitol, 88(4):822–825PubMedGoogle Scholar
  27. Wagner JT, Lüdemann H, Färber PM, Lottspeich F, Krauth-Siegel RL (1998) Glutamate dehydrogenase, the marker protein of Plasmodium falciparum, cloning, expression and characterization of the malarial enzyme. Eur J biochem, 258: 813–819CrossRefPubMedGoogle Scholar
  28. Yang Q, Mao WH, Ferre I, Bayon JE, Mao XZ, Gonzalez-Gallego J (1998) Plasma aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH) and gamma-glutamyl transpeptidase (γ-GTP) activities in water buffaloes with experimental subclinical fasciolosis. Vet Parasitol, 78(2):129–136CrossRefPubMedGoogle Scholar

Copyright information

© Indian Society for Parasitology 2009

Authors and Affiliations

  1. 1.Section of Parasitology, Department of ZoologyAligarh Muslim UniversityAligarhIndia

Personalised recommendations