Interaction of multidrug-resistant Chinese hamster ovary cells with the peptide ionophore gramicidin D

Abstract

A major form of multidrug resistance results from the overexpression of P-glycoprotein, a 170 kDa membrane protein. Multidrug resistant (MDR) Chinese hamster ovary (CHO) cells and mdrl transfectants displayed cross-resistance to the channel-forming peptide ionophore gramicidin D, which was reversed by various chemosensitizers, thus directly implicating P-glycoprotein as the mediator of resistance. However, gramicidin D was not able to inhibit [ 3H]azidopine photolabelling of P-glycoprotein. MDR cells were not resistant to other pore-forming ionophores, but showed a modest level of cross-resistance to the mobile ionophore valinomycin. There was no difference in 125I-gramicidin D uptake by resistant and sensitive cells. Resistant cells showed lower 86Rb + uptake, relative to the drug-sensitive parent. Addition of GmD increased both the rate and the level of 86Rb + uptake in sensitive cells, but had no effect on MDR cells. MDR cells also showed much lower rates of gramicidin D-dependent 86Rb + efflux than sensitive cells, and this was greatly increased by verapamil. These results suggest that P-glycoprotein interferes with the formation of ion-conducting gramicidin D channels. In contrast, valinomycin had the same effect on gramicidin D-dependent cation efflux in MDR and sensitive cells. Gramicidin D is thus unique among the ionophores in being a substrate for P-glycoprotein, which appears to greatly reduce the formation of active dimeric channels in the plasma membrane of MDR cells.