In archaebacteria, there is a doxorubicin efflux pump similar to mammalian P-glycoprotein.
We selected for study an anthracycline-resistant mutant from the archaebacteria Haloferax volcanii. This resistance was reversed by a Ca(2+)-channel antagonist, nifedipine (NDP). This resistance and its reversal by NDP suggest P-glycoprotein (Pgp) to be responsible for maintaining an anticancer drug concentration below the cytotoxic level. Using rhodamine 123 (RH123) as a substrate for Pgp, we then examined whether the resistance to anthracyclines in this bacteria might involve a Pgp-like anthracycline efflux pump. RH123 accumulation by the bacteria was determined with flow cytometry. A steady-state RH123 accumulation by the resistant cells revealed approx. one-fifteenth of that by the wild-type cells, which could be remarkably enhanced by NDP. The other modulators of Pgp, diltiazem and verapamil, also enhanced RH123 accumulation in resistant cells. The uncoupler FCCP completely restored RH123 accumulation in resistant cells to the wild-type cell level. RH123 unidirectional efflux from resistant cells after its preloading revealed much greater than that from wild-type cells, which was remarkably inhibited by FCCP. These confirmed that RH123 low accumulation involves its active efflux mechanism. Taken together, the present study indicated that lower evolutionary archaebacteria might also express a Pgp-like protein very similar to mammalian Pgp.
Department of Biophysics, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
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Miyauchi
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Komatsubara
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Kamo
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000818:Animals; D001105:Archaea / Q000187:drug effects / Q000378:metabolism*; D001412:Bacillus subtilis / Q000187:drug effects / Q000378:metabolism; D001693:Biological Transport, Active / Q000187:drug effects; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D004110:Diltiazem / Q000494:pharmacology; D004317:Doxorubicin / Q000378:metabolism* / Q000494:pharmacology; D004352:Drug Resistance, Microbial; D004357:Drug Synergism; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D008322:Mammals; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009543:Nifedipine / Q000494:pharmacology; D020112:Rhodamine 123; D012235:Rhodamines / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014700:Verapamil / Q000494:pharmacology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D005456:Fluorescent Dyes; D008562:Membrane Glycoproteins; D012235:Rhodamines; D020112:Rhodamine 123; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D004317:Doxorubicin; D014700:Verapamil; D004110:Diltiazem; D009543:Nifedipine