Enzymes of carbohydrate metabolism in Leishmania donovani amastigotes.
A method for the isolation of Leishmania donovani amastigotes from infected hamster spleen and liver tissues is described. Over 85% of the isolated amastigotes were viable as judged by acridine orange-ethidium bromide staining and in vitro transformation to the promastigote form. A comprehensive survey of the enzymes of carbohydrate metabolism in L. donovani amastigotes and promastigotes was conducted. Amastigotes and promastigotes possess all of the enzymes of the Embden-Meyerhof pathway, hexose monophosphate shunt, and tricarboxylic acid cycle. Cell-free extracts of both forms demonstrate an active glutamate dehydrogenase, thus linking activity which permits entry of pyruvate into the tricarboxylic acid cycle. Both forms demonstrate an active glutamate dehydrogenase, thus linking amino acid metabolism with carbohydrate metabolism. Pyruvate carboxylase, the enzyme responsible for replenishment of C4 acids by heterotrophic CO2 fixation into pyruvate, was also demonstrable in the tissue and insect forms. In general, activities of promastigote enzymes are higher than the amastigote enzymes. Differences between the vertebrate (amastigote) and invertebrate (promastigote) forms in their potential to utilize carbohydrates as substrates would appear to be quantitative rather than qualitative.
D000818:Animals; D050260:Carbohydrate Metabolism; D002952:Citric Acid Cycle; D006224:Cricetinae; D004734:Energy Metabolism; D005260:Female; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D006600:Hexosephosphates / Q000378:metabolism; D007891:Leishmania / Q000201:enzymology* / Q000302:isolation & purification; D007898:Leishmaniasis, Visceral / Q000469:parasitology; D008099:Liver / Q000469:parasitology; D008647:Mesocricetus; D010084:Oxidation-Reduction; D011766:Pyruvate Carboxylase / Q000378:metabolism; D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism; D013154:Spleen / Q000469:parasitology
D006600:Hexosephosphates; D011768:Pyruvate Dehydrogenase Complex; D005969:Glutamate Dehydrogenase; D011766:Pyruvate Carboxylase