A key role for old yellow enzyme in the metabolism of drugs by Trypanosoma cruzi.
Trypanosoma cruzi is the etiological agent of Chagas' disease. So far, first choice anti-chagasic drugs in use have been shown to have undesirable side effects in addition to the emergence of parasite resistance and the lack of prospect for vaccine against T. cruzi infection. Thus, the isolation and characterization of molecules essential in parasite metabolism of the anti-chagasic drugs are fundamental for the development of new strategies for rational drug design and/or the improvement of the current chemotherapy. While searching for a prostaglandin (PG) F(2alpha) synthase homologue, we have identified a novel "old yellow enzyme" from T. cruzi (TcOYE), cloned its cDNA, and overexpressed the recombinant enzyme. Here, we show that TcOYE reduced 9,11-endoperoxide PGH(2) to PGF(2alpha) as well as a variety of trypanocidal drugs. By electron spin resonance experiments, we found that TcOYE specifically catalyzed one-electron reduction of menadione and beta-lapachone to semiquinone-free radicals with concomitant generation of superoxide radical anions, while catalyzing solely the two-electron reduction of nifurtimox and 4-nitroquinoline-N-oxide drugs without free radical production. Interestingly, immunoprecipitation experiments revealed that anti-TcOYE polyclonal antibody abolished major reductase activities of the lysates toward these drugs, identifying TcOYE as a key drug-metabolizing enzyme by which quinone drugs have their mechanism of action.
Kubata
Bruno Kilunga
BK
Department of Molecular Behavioral Biology, Osaka Bioscience Institute, Osaka 565-0874, Japan. kubata@obi.or.jp
Kabututu
Zakayi
Z
Nozaki
Tomoyoshi
T
Munday
Craig J
CJ
Fukuzumi
Shunichi
S
Ohkubo
Kei
K
Lazarus
Michael
M
Maruyama
Toshihiko
T
Martin
Samuel K
SK
Duszenko
Michael
M
Urade
Yoshihiro
Y
D000595:Amino Acid Sequence; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism / Q000494:pharmacology; D001241:Aspirin / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D002384:Catalysis; D016861:Cyclooxygenase Inhibitors / Q000378:metabolism / Q000494:pharmacology; D016054:DNA, Protozoan; D015237:Dinoprost / Q000096:biosynthesis; D015232:Dinoprostone / Q000096:biosynthesis; D015870:Gene Expression; D006911:Hydroxyprostaglandin Dehydrogenases / Q000235:genetics; D007213:Indomethacin / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D009252:NADPH Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D009285:Naphthoquinones / Q000378:metabolism; D010084:Oxidation-Reduction; D015230:Prostaglandin D2 / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D014344:Trypanocidal Agents / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000235:genetics
D000894:Anti-Inflammatory Agents, Non-Steroidal; D016861:Cyclooxygenase Inhibitors; D016054:DNA, Protozoan; D009285:Naphthoquinones; D014344:Trypanocidal Agents; D015237:Dinoprost; D006911:Hydroxyprostaglandin Dehydrogenases; C031051:prostaglandin-F synthase; D009252:NADPH Dehydrogenase; D015232:Dinoprostone; D001241:Aspirin; D015230:Prostaglandin D2; D007213:Indomethacin