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

doi:10.1073/pnas.1203632109

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 journal › Article › peer-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 language	English
Pages (from-to)	12473-12478
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	109
Issue number	31
DOIs	https://doi.org/10.1073/pnas.1203632109
State	Published - 31 Jul 2012
Externally published	Yes

Keywords

Multidrug resistance
Secondary transport

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10.1073/pnas.1203632109

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title = "Manipulating the drug/proton antiport stoichiometry of the secondary multidrug transporter MdfA",

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.",

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T1 - Manipulating the drug/proton antiport stoichiometry of the secondary multidrug transporter MdfA

AU - Tirosh, Osnat

AU - Sigal, Nadejda

AU - Gelman, Amir

AU - Sahar, Nadav

AU - Fluman, Nir

AU - Siemion, Shira

AU - Bibi, Eitan

PY - 2012/7/31

Y1 - 2012/7/31

N2 - 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.

AB - 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.

KW - Multidrug resistance

KW - Secondary transport

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U2 - 10.1073/pnas.1203632109

DO - 10.1073/pnas.1203632109

M3 - مقالة

C2 - 22802625

SN - 0027-8424

VL - 109

SP - 12473

EP - 12478

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 31

ER -

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

Abstract

Keywords

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