Effect of DL-alpha-difluoromethylornithine on polyamine synthesis and interconversion in Trichomonas vaginalis grown in a semi-defined medium.
Growth of Trichomonas vaginalis in a semi-defined medium was inhibited by 5 mM DL-alpha-difluoromethylornithine (DFMO). Using high pressure liquid chromatography (HPLC) analysis, putrescine and cadaverine levels were found to be 90 and 100% reduced, respectively after 120 h exposure, whilst spermidine and spermine levels were unchanged. Putrescine (40 microM) and cadaverine (6 microM) were detected in the spent media from control cultures. Neither of these diamines was detected in spent media from 72 h DFMO-treated cultures. Changes in intracellular levels of amine precursors were also determined by HPLC. There was a transient increase in ornithine to 39 nmol (mg protein)-1 at 48 h in the DFMO-treated cells while it remained undetectable in control cells throughout the experiment. Arginine and citrulline levels remained high, decreasing to control levels only after 72 h. Only spermine (1 mM) rescued DFMO-treated cells, and this is discussed with respect to the presence of a putative spermine-specific oxidase designated by its sensitivity to aminoguanidine. Aerobic incubation of growing (normal) cells with [14C]spermine resulted in the production of an unknown metabolite (19% of total label), whose content was reduced to 5% under anaerobic conditions. Decarboxylated S-adenosylmethionine remained undetectable in DFMO-treated cells, and the methylation index (ratio of S-adenosylmethionine to S-adenosylhomocysteine) did not change from the control value of 9.3. Ornithine decarboxylase, S-adenosylmethionine synthetase, S-adenosylmethionine:L-homocysteine methyltransferase, and S-adenosylhomocysteine hydrolase enzyme activities were detected. However, S-adenosylmethionine decarboxylase, spermidine synthase or spermine synthase were not detected. These findings are discussed with reference to the arginine dihydrolase pathway whose end products are putrescine and ATP.(ABSTRACT TRUNCATED AT 250 WORDS)
D000241:Adenosine / Q000378:metabolism; D000818:Animals; D002103:Cadaverine / Q000096:biosynthesis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D000518:Eflornithine / Q000494:pharmacology*; D009955:Ornithine Decarboxylase / Q000378:metabolism; D011048:Polarography; D011073:Polyamines / Q000096:biosynthesis* / Q000378:metabolism; D011700:Putrescine / Q000096:biosynthesis / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism; D013095:Spermidine / Q000096:biosynthesis / Q000378:metabolism; D013096:Spermine / Q000096:biosynthesis / Q000378:metabolism; D014246:Trichomonas vaginalis / Q000187:drug effects* / Q000201:enzymology
D003470:Culture Media; D011073:Polyamines; D013096:Spermine; D012436:S-Adenosylmethionine; D009955:Ornithine Decarboxylase; D000241:Adenosine; D002103:Cadaverine; D013095:Spermidine; D011700:Putrescine; D000518:Eflornithine