Leishmania mexicana: subcellular distribution of enzymes in amastigotes and promastigotes.
Glycosomes and mitochondrial vesicles from cultured promastigotes of Leishmania mexicana mexicana have been separated using isopycnic centrifugation on linear sucrose gradients. Hexokinase (EC 2.7.1.2), glucose phosphate isomerase (EC 5.3.1.9), phosphofructokinase (EC 2.7.1.11), glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12), and phosphoenolpyruvate carboxykinase (EC 4.1.1.49) were recovered largely in association with glycosomes (density; 1.215 g/ml). Phosphoglycerate kinase (EC 2.7.2.3) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49) had some small glycosomal activity, but were mostly recovered in the soluble fractions. Malate dehydrogenase (EC 1.1.1.37) showed a broad peak corresponding to that of the mitochondrial marker oligomycin-sensitive ATPase (EC 3.6.1.4) (density; 1.190 g/ml). Glutamate dehydrogenase (EC 1.4.1.3) and alanine aminotransferase (EC 2.6.1.2) both showed small mitochondrial peaks, but most of the activities were recovered elsewhere on the gradient and in the soluble fractions. The subcellular location of enzymes in L.m. mexicana amastigotes was investigated by following the release of soluble enzymes from digitonin-treated amastigotes. This revealed distinct cytosolic, mitochondrial, and glycosomal compartments. The findings give an insight into the organization and control of L.m. mexicana promastigote and amastigote energy metabolism.
Mottram
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JC
Coombs
G H
GH
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000410:Alanine Transaminase / Q000378:metabolism; D004072:Digitonin / Q000494:pharmacology; D005956:Glucose-6-Phosphate Isomerase / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism; D007521:Isocitrate Dehydrogenase / Q000378:metabolism; D007891:Leishmania / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D008291:Malate Dehydrogenase / Q000378:metabolism; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000378:metabolism; D011770:Pyruvate Kinase / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology
D008291:Malate Dehydrogenase; D007521:Isocitrate Dehydrogenase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005969:Glutamate Dehydrogenase; D000410:Alanine Transaminase; D006593:Hexokinase; D011770:Pyruvate Kinase; D000251:Adenosine Triphosphatases; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D005956:Glucose-6-Phosphate Isomerase; D004072:Digitonin