Manipulating the drug/proton antiport stoichiometry of the secondary multidrug transporter MdfA

Osnat Tirosh, Nadejda Sigal, Amir Gelman, Nadav Sahar, Nir Fluman, Shira Siemion, Eitan Bibi

Research output: Contribution to journalArticlepeer-review

Abstract

Multidrug transporters are integral membrane proteins that use cellular energy to actively extrude antibiotics and other toxic compounds from cells. The multidrug/proton antiporter MdfA from Escherichia coli exchanges monovalent cationic substrates for protons with a stoichiometry of 1, meaning that it translocates only one proton per antiport cycle. This may explain why transport of divalent cationic drugs by MdfA is energetically unfavorable. Remarkably, however, we show that MdfA can be easily converted into a divalent cationic drug/≥2 proton-antiporter, either by random mutagenesis or by rational design. The results suggest that exchange of divalent cationic drugs with two (or more) protons requires an additional acidic residue in the multidrug recognition pocket of MdfA. This outcome further illustrates the exceptional promiscuous capabilities of multidrug transporters.

Original languageEnglish
Pages (from-to)12473-12478
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number31
DOIs
StatePublished - 31 Jul 2012

All Science Journal Classification (ASJC) codes

  • General

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