A New Twist in ABC Transporter Mediated Multidrug Resistance – Pdr5 is a Drug/proton Co-transporter

[Display omitted] •Pdr5 reconstituted in planar lipid bilayers displays ion channel activity, with a slight cation selectivity (PK+:PCl-=2.6:1).•ATP/Mg2+ and substrate are required to induce a membrane potential of Vm = 58 mV, corresponding to Vm = −58 mV in the cytosol in vivo.•Pdr5 co-transports H+ and substrate in a ATP/Mg2+-dependent manner.•H+ transport can be visualized in an in vitro transport assay. The two major efflux pump systems that are involved in multidrug resistance (MDR) are (i) ATP binding cassette (ABC) transporters and (ii) secondary transporters. While the former use binding and hydrolysis of ATP to facilitate export of cytotoxic compounds, the latter utilize electrochemical gradients to expel their substrates. Pdr5 from Saccharomyces cerevisiae is a prominent member of eukaryotic ATP binding cassette (ABC) transporters that are involved in multidrug resistance (MDR) and used as a frequently studied model system. Although investigated for decades, the underlying molecular mechanisms of drug transport and substrate specificity remain elusive. Here, we provide electrophysiological data on the reconstituted Pdr5 demonstrating that this MDR efflux pump does not only actively translocate its substrates across the lipid bilayer, but at the same time generates a proton motif force in the presence of Mg2+-ATP and substrates by acting as a proton/drug co-transporter. Importantly, a strictly substrate dependent co-transport of protons was also observed in in vitro transport studies using Pdr5-enriched plasma membranes. We conclude from these results that the mechanism of MDR conferred by Pdr5 and likely other transporters is more complex than the sole extrusion of cytotoxic compounds and involves secondary coupled processes suitable to increase the effectiveness.