Sensitivity of trypanosomatid protozoa to DFMO and metabolic turnover of ornithine decarboxylase

被引:28
作者
Carrillo, C
Cejas, S
Cortés, M
Ceriani, C
Huber, A
González, NS
Algranati, ID
机构
[1] Univ Buenos Aires, Fac Ciencias Exactas & Nat, Fdn Campomar, Inst Invest Bioquim, RA-1405 Buenos Aires, DF, Argentina
[2] Consejo Nacl Invest Cient & Tecn, RA-1405 Buenos Aires, DF, Argentina
关键词
ornithine decarboxylase; trypanosomatids; proliferation; polyamines; DFMO-resistance; Crithidia fasciculata; Phytomonas; Leishmania mexicana; Trypanosoma cruzi;
D O I
10.1006/bbrc.2000.3996
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
alpha -Difluoromethylornithine (DFMO), the specific and irreversible inhibitor of ornithine decarboxylase (ODC), was able to induce the arrest of proliferation in Leishmania mexicana and ODC-transformed Trypanosoma cruzi cultures grown in a semi-defined medium essentially free of polyamines. Conversely, Crithidia fasciculata and Phytomonas 274 were not affected by the inhibitor. The drug-resistance of Crithidia and Phytomonas was neither caused by an impairment of DFMO uptake nor by a decrease of the enzyme affinity for the inhibitor. We were also able to rule out the possibility of ODC overexpression in the drug-tolerant parasites. The measurements of ODC metabolic turnover indicated that the enzymes from Crithidia and Phytomonas have a short half-life of 20-40 min, while ODC from Leishmania and transgenic Trypanosoma cruzi are rather stable with a half-life longer than 6 hours. Analyses of polyamine internal pools under different growth conditions have shown that DFMO was able to markedly decrease the levels of putrescine and spermidine in all parasites, but the depletion of spermidine was higher in trypanosomatids containing an ODC with slow turnover. Our results suggest that in these parasites cultivated in the presence of the drug, spermidine might decrease below critical levels needed to maintain trypanothione concentrations or other conditions essential for normal proliferation. (C) 2000 Academic Press.
引用
收藏
页码:663 / 668
页数:6
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