Switch loop flexibility affects substrate transport of the AcrB efflux pump

Muller, Reinke T.; Travers, Timothy; Cha, Hi-jea; Phillips, Joshua L.; Gnanakaran, Sandrasegaram; Pos, Klaas M.

doi:10.1016/j.jmb.2017.09.018

Title: Switch loop flexibility affects substrate transport of the AcrB efflux pump

Journal Article · Thu Oct 05 00:00:00 EDT 2017 · Journal of Molecular Biology

DOI:https://doi.org/10.1016/j.jmb.2017.09.018· OSTI ID:1402663

Muller, Reinke T. ^[1];

^[2]; Cha, Hi-jea ^[3]; Phillips, Joshua L. ^[4];

^[2]; Pos, Klaas M. ^[1]

Goethe Univ. Frankfurt, Frankfurt am Main (Germany)
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Goethe Univ. Frankfurt, Frankfurt am Main (Germany); Engelhard Arzneimittel GmbH & Co. KG, Niederdorfelden (Germany)
Middle Tennessee State Univ., Murfreesboro, TN (United States)

The functionally important switch-loop of the trimeric multidrug transporter AcrB separates the access and deep drug binding pockets in every protomer. This loop, comprising 11 amino acid residues, has been shown to be crucial for substrate transport, as drugs have to travel past the loop to reach the deep binding pocket and from there are transported outside the cell via the connected AcrA and TolC channels. It contains four symmetrically arranged glycine residues suggesting that flexibility is a key feature for pump activity. Upon combinatorial substitution of these glycine residues to proline, functional and structural asymmetry was observed. Proline substitutions on the PC1 proximal side completely abolished transport and reduced backbone flexibility of the switch loop, which adopted a conformation restricting the pathway towards the deep binding pocket. Here, two phenylalanine residues located adjacent to the substitution sensitive glycine residues play a role in blocking the pathway upon rigidification of the loop, since the removal of the phenyl rings from the rigid loop restores drug transport activity.

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Research Organization:: Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

Sponsoring Organization:: USDOE Laboratory Directed Research and Development (LDRD) Program

Grant/Contract Number:: AC52-06NA25396

OSTI ID:: 1402663

Alternate ID(s):: OSTI ID: 1549482

Report Number(s):: LA-UR-17-28721; TRN: US1703120

Journal Information:: Journal of Molecular Biology, Vol. 429, Issue 24; ISSN 0022-2836

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 16 works

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