title: Functional expression of a multidrug P-glycoprotein transporter of Leishmania.
abstract: 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.
authors.affiliation: 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.
authors.forename: Fernando
authors.initials: F
authors.lastname: Cortés-Selva
authors.forename: Francisco
authors.initials: F
authors.lastname: Muñoz-Martínez
authors.forename: Attila
authors.initials: A
authors.lastname: Iliás
authors.forename: Antonio Ignacio
authors.initials: AI
authors.lastname: Jiménez
authors.forename: András
authors.initials: A
authors.lastname: Váradi
authors.forename: Francisco
authors.initials: F
authors.lastname: Gamarro
authors.forename: Santiago
authors.initials: S
authors.lastname: Castanys
mesh_terms: 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
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D001562:Benzimidazoles; D012717:Sesquiterpenes; C017807:bisbenzimide ethoxide trihydrochloride