Differential energetic metabolism during Trypanosoma cruzi differentiation. II. Hexokinase, phosphofructokinase and pyruvate kinase.
The activities of hexokinase (ATP:hexose-6-phosphate transferase, E.C. 2.7.1.1), phosphofructokinase (ATP:fructose-6-phosphate 1-phosphotransferase, E.C.2.7.1.11) and pyruvate kinase (ATP:pyruvate transferase, E.C. 2.7.1.40), and their kinetic behaviour in two morphological forms of Trypanosoma cruzi (epimastigotes and metacyclic trypomastigotes) have been studied. The kinetic responses of the three enzymes to their respective substrates were normalized to hyperbolic forms on a velocity versus substrate concentration plots. Hexokinase and phosphofructokinase showed a higher activity in epimastigotes than in metacyclics, whereas pyruvate kinase had similar activity in both forms of the parasite. The specific activity of hexokinase from epimastigotes was 102.00 mUnits/mg of protein and the apparent Km value for glucose was 35.4 microM. Metacyclic forms showed a specific activity of 55.25 mUnits/mg and a Km value of 46.3 microM. The kinetic parameters (specific activity and Km for fructose 6-phosphate) of phosphofructokinase for epimastigotes were 42.60 mUnits/mg and 0.31 mM and for metacyclics 13.97 mUnits/mg and 0.16 mM, respectively. On the contrary, pyruvate kinase in both forms of T. cruzi did not show significant differences in its kinetic parameters. The specific activity in epimastigotes was 37.00 mUnits/mg and the Km for phosphoenolpyruvate was 0.47 mM, whereas in metacyclics these values were 42.94 mUnits/mg and 0.46 mM, respectively. The results presented in this work, clearly demonstrate a quantitative change in the glycolytic pathway of both culture forms of T. cruzi.
Adroher
F J
FJ
Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Spain.
Osuna
A
A
Lupiáñez
J A
JA
D000818:Animals; D002454:Cell Differentiation; D004734:Energy Metabolism; D005636:Fructosephosphates / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006593:Hexokinase / Q000378:metabolism*; D007700:Kinetics; D010728:Phosphoenolpyruvate / Q000494:pharmacology; D010732:Phosphofructokinase-1 / Q000378:metabolism*; D011770:Pyruvate Kinase / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D013379:Substrate Specificity; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000254:growth & development*
D005636:Fructosephosphates; D000644:Quaternary Ammonium Compounds; D010728:Phosphoenolpyruvate; D006593:Hexokinase; D010732:Phosphofructokinase-1; D011770:Pyruvate Kinase; D005947:Glucose