Differential energetic metabolism during Trypanosoma cruzi differentiation. I. Citrate synthase, NADP-isocitrate dehydrogenase, and succinate dehydrogenase.
The activities of the mitochondrial enzymes citrate synthase (citrate oxaloacetatelyase, EC 4.1.3.7), NADP-linked isocitrate dehydrogenase (threo-Ds-isocitrate:NADP+ oxidoreductase (decarboxylating), EC 1.1.1.42), and succinate dehydrogenase (succinate: FAD oxidoreductase, EC 1.3.99.1) as well as their kinetic behavior in the two developmental forms of Trypanosoma cruzi at insect vector stage, epimastigotes and infective metacyclic trypomastigotes, were studied. The results presented in this work clearly demonstrate a higher mitochondrial metabolism in the metacyclic forms as is shown by the extraordinary enhanced activities of metacyclic citrate synthase, isocitrate dehydrogenase, and succinate dehydrogenase. In epimastigotes, the specific activities of citrate synthase at variable concentrations of oxalacetate and acetyl-CoA were 24.6 and 26.6 mU/mg of protein, respectively, and the Michaelis constants were 7.88 and 6.84 microM for both substrates. The metacyclic enzyme exhibited the following kinetic parameters: a specific activity of 228.4 mU/mg and Km of 3.18 microM for oxalacetate and 248.5 mU/mg and 2.75 microM, respectively, for acetyl-CoA. NADP-linked isocitrate dehydrogenase specific activities for epimastigotes and metacyclics were 110.2 and 210.3 mU/mg, whereas the apparent Km's were 47.9 and 12.5 microM, respectively. No activity for the NAD-dependent isozyme was found in any form of T. cruzi differentiation. The particulated succinate dehydrogenase showed specific activities of 8.2 and 39.1 mU/mg for epimastigotes and metacyclic trypomastigotes, respectively, although no significant changes in the Km (0.46 and 0.48 mM) were found. The cellular role and the molecular mechanism that probably take place during this significant shift in the mitochondrial metabolism during the T. cruzi differentiation have been discussed.
Adroher
F J
FJ
Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Spain.
Osuna
A
A
Lupiañez
J A
JA
D000818:Animals; D050260:Carbohydrate Metabolism; D002454:Cell Differentiation; D002950:Citrate (si)-Synthase / Q000378:metabolism*; D004734:Energy Metabolism; D007521:Isocitrate Dehydrogenase / Q000378:metabolism*; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology; D007652:Oxo-Acid-Lyases / Q000378:metabolism*; D013385:Succinate Dehydrogenase / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000254:growth & development
D007521:Isocitrate Dehydrogenase; C039355:isocitrate dehydrogenase (NADP+); D013385:Succinate Dehydrogenase; D002950:Citrate (si)-Synthase; D007652:Oxo-Acid-Lyases