The Leishmania ATP-binding cassette protein PGPA is an intracellular metal-thiol transporter ATPase.
The Leishmania ATP-binding cassette (ABC) transporter PGPA is involved in metal resistance (arsenicals and antimony), although the exact mechanism by which PGPA confers resistance to antimony, the first line drug against Leishmania, is unknown. The results of co-transfection experiments, transport assays, and the use of inhibitors suggest that PGPA recognizes metals conjugated to glutathione or trypanothione, a glutathione-spermidine conjugate present in Leishmania. The HA epitope tag of the influenza hemagglutinin as well as the green fluorescent protein were fused at the COOH terminus of PGPA. Immunofluorescence, confocal, and electron microscopy studies of the fully functional tagged molecules clearly indicated that PGPA is localized in membranes that are close to the flagellar pocket, the site of endocytosis and exocytosis in this parasite. Subcellular fractionation of Leishmania tarentolae PGPAHA transfectants was performed to further characterize this ABC transporter. The basal PGPA ATPase activity was determined to be 115 nmol/mg/min. Transport experiments using radioactive arsenite-glutathione conjugates clearly showed that PGPA recognizes and actively transports thiol-metal conjugates. Overall, the results are consistent with PGPA being an intracellular ABC transporter that confers arsenite and antimonite resistance by sequestration of the metal-thiol conjugates.
Légaré
D
D
Centre de Recherche en Infectiologie du Centre Hospitalier de l'Université Laval, 2705 boulevard Laurier Ste-Foy, Québec G1V 4G2, Canada.
Richard
D
D
Mukhopadhyay
R
R
Stierhof
Y D
YD
Rosen
B P
BP
Haimeur
A
A
Papadopoulou
B
B
Ouellette
M
M
D018528:ATP-Binding Cassette Transporters / Q000032:analysis / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D007891:Leishmania; D008562:Membrane Glycoproteins / Q000032:analysis / Q000378:metabolism*; D008670:Metals / Q000378:metabolism; D015800:Protozoan Proteins; D013438:Sulfhydryl Compounds / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D008562:Membrane Glycoproteins; D008670:Metals; C106891:PgpA protein, Leishmania; D015800:Protozoan Proteins; D013438:Sulfhydryl Compounds; D000251:Adenosine Triphosphatases