Functional role of phosphorylation of the multidrug transporter (P-glycoprotein) by protein kinase C in multidrug-resistant MCF-7 cells.
Multidrug resistance (MDR) is the phenomenon in which cells become resistant to several classes of structurally and functionally diverse drugs after exposure to a single cytotoxic agent. One form of MDR is associated with the overexpression of a large plasma membrane phosphoglycoprotein, P-glycoprotein, which acts as an energy-requiring drug transport pump. Protein kinase C may participate in MDR through posttranslational modification of P-glycoprotein. The purpose of this study is to critically evaluate P-glycoprotein as a substrate for protein kinase C and to determine whether phosphorylation leads to changes in drug transport. Protein kinase C from rat brain phosphorylated immunoprecipitated P-glycoprotein in a manner dependent on the activation of the exogenous kinase. Phorbol 12-myristate 13-acetate (PMA) increased the phosphorylation of P-glycoprotein 6-fold and selectively decreased the accumulation of vinblastine in resistant MCF-7/AdrR cells. PMA selectively decreased the cellular association of vinblastine with MDR cells after brief periods of incubation, but only after critical concentrations of drug were achieved. The actions of PMA did not require new synthesis of P-glycoprotein. PMA had similar effects in MCF-7/BC-19, a cell line transfected with a cDNA for P-glycoprotein. Staurosporine inhibited the effects of PMA on the phosphorylation of P-glycoprotein and on the accumulation of vinblastine. These data demonstrated that immunoprecipitated P-glycoprotein can be a substrate for protein kinase C, and that phosphorylation of the transporter is associated with significant changes in drug transport.
Aftab
D T
DT
Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510.
Yang
J M
JM
Hait
W N
WN
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D003513:Cycloheximide / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D004317:Doxorubicin / Q000494:pharmacology; D018432:Drug Resistance, Multiple / Q000502:physiology*; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured; D014747:Vinblastine / Q000493:pharmacokinetics
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000470:Alkaloids; D018076:DNA, Complementary; D014747:Vinblastine; D004317:Doxorubicin; D003513:Cycloheximide; D011493:Protein Kinase C; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate