P-glycoprotein-like protein contributes to cadmium resistance in Euglena gracilis.
Selective pressures from polluted environments have led to the development of resistance systems in aquatic organisms. Using different techniques, this study examined a cadmium defense mechanism of the freshwater unicellular protozoa Euglena gracilis, and found it to be an efflux pump similar to the multidrug resistance P-glycoprotein. Cd(2+)-treated E. gracilis were able to extrude Rhodamine 123 at 21 degrees C, but not at 4 degrees C. Furthermore, verapamil, a P-glycoprotein modulator, partially blocked the efflux process (at 21 degrees C), and enhanced the Cd(2+) toxic effects on these cells. Western immunoblots of cell lysates, using the anti-P-glycoprotein antibody JSB-1, revealed a 120-KDa protein, which was expressed, in high amounts on Cd(2+)-exposed cells (74% above the control values). Moreover, cells treated with JSB-1 became more sensitive to the harmful effects of cadmium, showing a decreased survival rate. Taken together, these results suggest that a MDR phenotype has evolved in Euglena as one of the mechanisms for cadmium detoxification.
Einicker-Lamas
M
M
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, 21941-590, Rio de Janeiro, Brazil. einicker@biof.ufrj.br
Morales
M M
MM
Miranda
K
K
Garcia-Abreu
J
J
Oliveira
A J F
AJ
Silva
F L S
FL
Oliveira
M M
MM
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000032:analysis / Q000276:immunology / Q000502:physiology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015153:Blotting, Western; D002104:Cadmium / Q000494:pharmacology* / Q000633:toxicity; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance; D005056:Euglena gracilis / Q000187:drug effects / Q000502:physiology*; D026901:Membrane Transport Proteins / Q000502:physiology; D008856:Microscopy, Fluorescence; D020112:Rhodamine 123 / Q000378:metabolism / Q000494:pharmacology; D013696:Temperature; D014700:Verapamil / Q000494:pharmacology
D018435:ATP Binding Cassette Transporter, Subfamily B; D000911:Antibodies, Monoclonal; D026901:Membrane Transport Proteins; D002104:Cadmium; D020112:Rhodamine 123; D014700:Verapamil