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Decrease of P-Glycoprotein Activity in K562/ADR Cells by MβCD and Filipin and Lack of Effect Induced by Cholesterol Oxidase Indicate That This Transporter Is Not Located in Rafts

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Abstract

The effect of low-density membrane domains on function of the plasma membrane transporter P-glycoprotéine (P-gp), involved in multidrug resistance (MDR) phenotype, has been investigated in K562/ADR cells. To this end we reversibly altered the cholesterol content of K562/ADR cells by using methyl-β-cyclodextrin as a cholesterol chelator and conversely we repleted them through incubation with cholesterol in culture medium. We also used the cholesterol-binding fluorochrome filipin and cholesterol oxidase. Our data show that either cholesterol depletion or complex formation with filipin resulted in a strong decrease of P-gp activity. However, when cells were incubated with cholesterol oxidase that are known to disrupt rafts, no modification of the P-gp activity was observed. In addition, using a free-detergent methodology to separate by ultracentrifugation, “light,” “heavy,” and “extra heavy” fractions we show that no P-gp is found in the “light” fraction where rafts are usually detected. Altogether, our data strongly suggest that, in this cell line, P-gp is not localized in rafts.

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Authors and Affiliations

  1. Laboratoire de Physicochimie Biomoléculaire et Cellulaire (LPBC/CSSB UMR 7033), Université Paris Nord, 74 rue Marcel Cachin, 93017, Bobigny, France

    Paiboon Reungpatthanaphong, Carole Marbeuf-Gueye, Laurence Le Moyec, Milena Salerno & Arlette Garnier-Suillerot

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  1. Paiboon Reungpatthanaphong
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  2. Carole Marbeuf-Gueye
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  3. Laurence Le Moyec
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  4. Milena Salerno
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  5. Arlette Garnier-Suillerot
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Correspondence to Carole Marbeuf-Gueye.

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Reungpatthanaphong, P., Marbeuf-Gueye, C., Moyec, L.L. et al. Decrease of P-Glycoprotein Activity in K562/ADR Cells by MβCD and Filipin and Lack of Effect Induced by Cholesterol Oxidase Indicate That This Transporter Is Not Located in Rafts. J Bioenerg Biomembr 36, 533–543 (2004). https://doi.org/10.1007/s10863-004-9000-8

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