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PLoS One. 2011;6(6):e21196. doi: 10.1371/journal.pone.0021196. Epub 2011 Jun 15.

Determination of real-time efflux phenotypes in Escherichia coli AcrB binding pocket phenylalanine mutants using a 1,2'-dinaphthylamine efflux assay.

Author information

1
Department of Medicine, Center for Infectious Diseases and Travel Medicine, Albert-Ludwigs-University, University Hospital, Freiburg, Germany. juergen@bohnert.name

Abstract

To evaluate the importance of phenylalanine residues for substrate transport in the Escherichia coli efflux pump protein AcrB, we subjected Phe-to-Ala binding pocket mutants to a real-time efflux assay with the novel near-infrared lipophilic membrane probe 1,2'-dinaphthylamine (1,2'-DNA). All mutations, with the exception of F617A, led to considerable retardation of efflux. F610A was the point mutation with the most pronounced impact, followed by F628A, F615A, F136A, and F178A. This is the first study to demonstrate the importance of single phenylalanine residues within the AcrB binding pocket for real-time substrate transport.

PMID:
21698261
PMCID:
PMC3115983
DOI:
10.1371/journal.pone.0021196
[Indexed for MEDLINE]
Free PMC Article

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