Arginine decarboxylase in Trypanosoma cruzi, characteristics and kinetic properties.
Polyamines are known to play an essential role in cell growth and differentiation. In animals, putrescine is mainly synthesized from ornithine by ornithine decarboxylase (ODC). In higher plants and in bacteria putrescine can also be synthesized from arginine by arginine decarboxylase (ADC). In this paper we report the presence of significant levels of ADC activity in crude extracts of Trypanosoma cruzi, RA strain epimastigotes. ADC activity was detected during a very narrow time range, corresponding to the early logarithmic growth phase. This activity was inhibited by DL-alpha-difluoromethylarginine, a specific irreversible inhibitor of ADC and activated by DL-alpha-difluoromethylornithine, a specific irreversible inhibitor of ODC. The reaction showed an absolute requirement for pyridoxal phosphate, dithiothreitol and Mg++. The enzyme half life was about 10 hrs., showed maximum activity at pH 7.9 and a Km for arginine of 5 mM. ADC activity was stimulated by fetal-calf-serum and inhibited by spermine, probably through a negative feed-back regulation on the levels of the enzyme. ODC activity was not detected. These results confirm our previous reports on the capability of T. cruzi, RA strain epimastigotes to synthesize putrescine from arginine via agmatine by ADC and point out differences on polyamine metabolism between the parasite and the mammalian host cell.
Hernández
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Departamento de Bioquímica Humana, Facultad de Medicina, Universidad de Buenos Aires, Republica Argentina.
Schwarcz de Tarlovsky
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D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003470:Culture Media; D000518:Eflornithine; D004789:Enzyme Activation; D004791:Enzyme Inhibitors; D007700:Kinetics; D009955:Ornithine Decarboxylase / Q000378:metabolism; D011073:Polyamines; D012710:Serum Albumin, Bovine; D014349:Trypanosoma cruzi / Q000201:enzymology*
D003470:Culture Media; D004791:Enzyme Inhibitors; D011073:Polyamines; D012710:Serum Albumin, Bovine; C030965:alpha-(difluoromethyl)arginine; D001120:Arginine; D002262:Carboxy-Lyases; D009955:Ornithine Decarboxylase; C019722:arginine decarboxylase; D000518:Eflornithine