Shukla Suneet, Sauna Zuben E, Prasad Rajendra, Ambudkar Suresh V
Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4256, USA.
Biochem Biophys Res Commun. 2004 Sep 17;322(2):520-5. doi: 10.1016/j.bbrc.2004.07.151.
To find novel drugs for effective antifungal therapy in candidiasis, we examined disulfiram, a drug used for the treatment of alcoholism, for its role as a potential modulator of Candida multidrug transporter Cdr1p. We show that disulfiram inhibits the oligomycin-sensitive ATPase activity of Cdr1p and 2.5mM dithiothreitol reverses this inhibition. Disulfiram inhibited the binding of photoaffinity analogs of both ATP ([alpha-(32)P]8-azidoATP; IC(50)=0.76 microM) and drug-substrates ([(3)H]azidopine and [(125)I]iodoarylazidoprazosin; IC(50) approximately 12 microM) to Cdr1p in a concentration-dependent manner, suggesting that it can interact with both ATP and substrate-binding site(s) of Cdr1p. Furthermore, a non-toxic concentration of disulfiram (1 microM) increased the sensitivity of Cdr1p expressing Saccharomyces cerevisiae cells to antifungal agents (fluconazole, miconazole, nystatin, and cycloheximide). Collectively these results demonstrate that disulfiram reverses Cdr1p-mediated drug resistance by interaction with both ATP and substrate-binding sites of the transporter and may be useful for antifungal therapy.
为了寻找用于念珠菌病有效抗真菌治疗的新型药物，我们研究了用于治疗酒精中毒的双硫仑作为白色念珠菌多药转运蛋白Cdr1p潜在调节剂的作用。我们发现双硫仑抑制Cdr1p的寡霉素敏感ATP酶活性，2.5mM二硫苏糖醇可逆转这种抑制作用。双硫仑以浓度依赖性方式抑制ATP（[α-(32)P]8-叠氮基ATP；IC(50)=0.76 microM）和药物底物（[(3)H]叠氮平及[(125)I]碘芳基叠氮哌唑嗪；IC(50)约为12 microM）的光亲和类似物与Cdr1p的结合，这表明它可与Cdr1p的ATP和底物结合位点相互作用。此外，无毒浓度的双硫仑（1 microM）增加了表达Cdr1p的酿酒酵母细胞对抗真菌剂（氟康唑、咪康唑、制霉菌素和环己酰亚胺）的敏感性。这些结果共同表明，双硫仑通过与转运蛋白的ATP和底物结合位点相互作用来逆转Cdr1p介导的耐药性，可能对抗真菌治疗有用。
