Purification and properties of phosphoglucose isomerases of Trypanosoma cruzi.
Glucosephosphate isomerase (PGI; EC 5.3.1.9) of Trypanosoma cruzi epimastigotes was found in about the same proportion in the glycosome and the cytosol. This subcellular distribution is similar to that of Leishmania mexicana, but contrasts with that of T. brucei bloodstream form, where the enzyme is essentially restricted to the glycosome. Glucosephosphate isomerase was highly purified from a glycosome-enriched fraction and to about 70% purity from the soluble extract. Both enzymes displayed Michaelis-Menten-Henri kinetics. Km values for fructose 6-phosphate were 0.125 +/- 0.07 and 0.80 +/- 0.10 mM for the glycosomal and the cytosolic PGIs, respectively. Erythrose-4-phosphate, 6-phosphogluconate and mannose-6-phosphate were inhibitors for both PGIs. Phosphogluconate and erythrose phosphate showed higher affinity for cytosolic PGI than for glycosomal PGI, by 2.5- and 4-fold respectively. The PGIs differed slightly in their isoelectric point (7.1 +/- 0.15 and 7.5 +/- 0.12) and optimum pH range. Both PGIs also differed in their chromatographic properties (ion-exchange and phenyl Sepharose), indicating a difference in charge and hydrophobicity, with the glycosomal enzyme being more hydrophobic. The molecular mass of both PGIs was 186,000 +/- 9000 Da, which is higher than that of other known PGIs, including those from T. brucei and other trypanosomatids. The molecular mass of the subunit, 63 kDa, is similar to that of PGIs from other sources. It appears that PGIs from T. cruzi are trimeric, in contrast with all other known PGIs which are dimeric.
D000818:Animals; D015153:Blotting, Western; D003600:Cytosol / Q000201:enzymology; D004072:Digitonin / Q000737:chemistry / Q000494:pharmacology; D005956:Glucose-6-Phosphate Isomerase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D007526:Isoelectric Point; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008970:Molecular Weight; D015388:Organelles / Q000201:enzymology; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013347:Subcellular Fractions; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000201:enzymology*