Cellular location and function of the P-glycoproteins (EhPgps) in Entamoeba histolytica multidrug-resistant trophozoites

Cecilia Bañuelos, Esther Orozco, Consuelo Gómez, Arturo González, Olivia Medel, Leobardo Mendoza, D. Guillermo Pérez

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

We have studied the cellular location and the efflux pump function of the Entamoeba histolytica P-glycoproteins (EhPgps) in drug-sensitive and -resistant trophozoites. Polyclonal antibodies against the EhPgp384 polypeptide (375-759 amino acids) revealed a 147-kDa protein by Western blot. The band intensity correlated with the emetine-resistance of the trophozoites. Through the confocal microscope, using the anti-EhPgp384 and fluorescein secondary antibodies, the EhPgps were found in a complex vesicular network, in the plasma membrane and outside of the cells. Transmission electron microscopy assays confirmed that drug-resistant trophozoites presented four to five times more EhPgps than sensitive cells. Fluorescence co-localization experiments using rhodamine-123 (R123) and the anti-EhPgp384 antibodies suggested the interaction between EhPgps and the drug. R123 efflux kinetics evidenced that the emetine-resistant trophozoites displayed a drug efflux kinetic four times higher than the drug-sensitive trophozoites, which was reduced by verapamil in both cases. EhPgps may participate in avoiding drug accumulation in the trophozoites by two putative mechanisms: (1) the direct extrusion of the drug from the plasma membrane, and (2) an indirect transport mechanism in which the drug is trapped by EhPgps and concentrated within vesicles that drive the drug to the plasma membrane.
Original languageAmerican English
Pages (from-to)291-300
Number of pages260
JournalMicrobial Drug Resistance
DOIs
StatePublished - 1 Jan 2002

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P-Glycoproteins
Trophozoites
Entamoeba histolytica
Pharmaceutical Preparations
Emetine
Rhodamine 123
Cell Membrane
Antibodies
Verapamil
Fluorescein
Transmission Electron Microscopy
Anti-Idiotypic Antibodies
Pharmacokinetics
Fluorescence
Western Blotting
Amino Acids
Peptides

Cite this

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title = "Cellular location and function of the P-glycoproteins (EhPgps) in Entamoeba histolytica multidrug-resistant trophozoites",
abstract = "We have studied the cellular location and the efflux pump function of the Entamoeba histolytica P-glycoproteins (EhPgps) in drug-sensitive and -resistant trophozoites. Polyclonal antibodies against the EhPgp384 polypeptide (375-759 amino acids) revealed a 147-kDa protein by Western blot. The band intensity correlated with the emetine-resistance of the trophozoites. Through the confocal microscope, using the anti-EhPgp384 and fluorescein secondary antibodies, the EhPgps were found in a complex vesicular network, in the plasma membrane and outside of the cells. Transmission electron microscopy assays confirmed that drug-resistant trophozoites presented four to five times more EhPgps than sensitive cells. Fluorescence co-localization experiments using rhodamine-123 (R123) and the anti-EhPgp384 antibodies suggested the interaction between EhPgps and the drug. R123 efflux kinetics evidenced that the emetine-resistant trophozoites displayed a drug efflux kinetic four times higher than the drug-sensitive trophozoites, which was reduced by verapamil in both cases. EhPgps may participate in avoiding drug accumulation in the trophozoites by two putative mechanisms: (1) the direct extrusion of the drug from the plasma membrane, and (2) an indirect transport mechanism in which the drug is trapped by EhPgps and concentrated within vesicles that drive the drug to the plasma membrane.",
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Cellular location and function of the P-glycoproteins (EhPgps) in Entamoeba histolytica multidrug-resistant trophozoites. / Bañuelos, Cecilia; Orozco, Esther; Gómez, Consuelo; González, Arturo; Medel, Olivia; Mendoza, Leobardo; Pérez, D. Guillermo.

In: Microbial Drug Resistance, 01.01.2002, p. 291-300.

Research output: Contribution to journalArticle

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T1 - Cellular location and function of the P-glycoproteins (EhPgps) in Entamoeba histolytica multidrug-resistant trophozoites

AU - Bañuelos, Cecilia

AU - Orozco, Esther

AU - Gómez, Consuelo

AU - González, Arturo

AU - Medel, Olivia

AU - Mendoza, Leobardo

AU - Pérez, D. Guillermo

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