Disulfiram is a potent modulator of multidrug transporter Cdr1p of Candida albicans.
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.
Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4256, USA.
Suneet
S
Shukla
Zuben E
ZE
Sauna
Rajendra
R
Prasad
Suresh V
SV
Ambudkar
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D001386:Azides / Q000378:metabolism; D002176:Candida albicans / Q000187:drug effects; D002177:Candidiasis / Q000188:drug therapy*; D003001:Cloning, Molecular; D004221:Disulfiram / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005656:Fungal Proteins / Q000187:drug effects* / Q000378:metabolism; D017930:Genes, Reporter; D026901:Membrane Transport Proteins / Q000187:drug effects* / Q000378:metabolism; D010761:Phosphorus Radioisotopes / Q000378:metabolism; D012441:Saccharomyces cerevisiae
D001386:Azides; C094693:CDR1 protein, Candida albicans; D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D010761:Phosphorus Radioisotopes; C017040:8-azidoadenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D004221:Disulfiram