Transforming a Drug/H+ Antiporter into a Polyamine Importer by a Single Mutation

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2012 Oct 5

PMID 23035252

Citations 24

Authors

Shlomo Brill

Ofir Sade Falk

Shimon Schuldiner

Affiliations

Soon will be listed here.

Abstract

EmrE, a multidrug antiporter from Escherichia coli, has presented biochemists with unusual surprises. Here we describe the transformation of EmrE, a drug/H(+) antiporter to a polyamine importer by a single mutation. Antibiotic resistance in microorganisms may arise by mutations at certain chromosomal loci. To investigate this phenomenon, we used directed evolution of EmrE to assess the rate of development of novel specificities in existing multidrug transporters. Strikingly, when a library of random mutants of EmrE was screened for resistance to two major antibacterial drugs--norfloxacin, a fluoroquinolone, and erythromycin, a macrolide--proteins with single mutations were found capable of conferring resistance. The mutation conferring erythromycin resistance resulted from substitution of a fully conserved and essential tryptophan residue to glycine, and, as expected, this protein lost its ability to recognize and transport the classical EmrE substrates. However, this protein functions now as an electrochemical potential driven importer of a new set of substrates: aliphatic polyamines. This mutant provides a unique paradigm to understand the function and evolution of distinct modes of transport.

Citing Articles

Dynamics underlie the drug recognition mechanism by the efflux transporter EmrE.

Li J, Her A, Besch A, Ramirez-Cordero B, Crames M, Banigan J Nat Commun. 2024; 15(1):4537.

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Functional promiscuity of small multidrug resistance transporters from Staphylococcus aureus, Pseudomonas aeruginosa, and Francisella tularensis.

Spreacker P, Wegrzynowicz A, Porter C, Beeninga W, Demas S, Powers E Mol Microbiol. 2024; 121(4):798-813.

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Activating alternative transport modes in a multidrug resistance efflux pump to confer chemical susceptibility.

Spreacker P, Thomas N, Beeninga W, Brousseau M, Porter C, Hibbs K Nat Commun. 2022; 13(1):7655.

PMID: 36496486 PMC: 9741644. DOI: 10.1038/s41467-022-35410-2.

High-pH structure of EmrE reveals the mechanism of proton-coupled substrate transport.

Shcherbakov A, Spreacker P, Dregni A, Henzler-Wildman K, Hong M Nat Commun. 2022; 13(1):991.

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Biologia futura: the role of polyamine in plant science.

Kamiab F, Tavassolian I, Hosseinifarahi M Biol Futur. 2021; 71(3):183-194.

PMID: 34554509 DOI: 10.1007/s42977-020-00027-3.

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