Functional expression of a multidrug P-glycoprotein transporter of Leishmania.
P-glycoprotein (Pgp) transporters play an important role in multidrug resistance in eukaryotic cells and in protozoan parasites such as Leishmania. To search for new reversal agents of the Leishmania tropica Pgp, we developed a screening assay using the Baculovirus-insect cell expression system. We demonstrated a MgATP-dependent, vanadate-sensitive transport of Hoechst 33342 in membrane preparations of Sf9 insect cells expressing Pgp. We have found that dihydro-beta-agarofuran sesquiterpenes from Maytenus cuzcoina inhibited Hoechst 33342 transport that correlates with their reversal effect in a multidrug-resistant L. tropica line overexpressing Pgp. The results suggest that Sf9 cell membrane Hoechst 33342 transport system represents an efficient tool for examining the interactions of Leishmania Pgp with pharmacological agents.
Cortés-Selva
Fernando
F
Instituto de Parasitología y Biomedicina López-Neyra, Consejo Superior de Investigaciones Científicas, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento s/n, 18100 Armilla, Granada, Spain.
Muñoz-Martínez
Francisco
F
Iliás
Attila
A
Jiménez
Antonio Ignacio
AI
Váradi
András
A
Gamarro
Francisco
F
Castanys
Santiago
S
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001562:Benzimidazoles / Q000493:pharmacokinetics*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D018432:Drug Resistance, Multiple / Q000502:physiology*; D007891:Leishmania / Q000187:drug effects / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D012717:Sesquiterpenes / Q000494:pharmacology; D018411:Spodoptera / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D001562:Benzimidazoles; D012717:Sesquiterpenes; C017807:bisbenzimide ethoxide trihydrochloride