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Conformational dynamics of the ABC transporter McjD seen by single-molecule FRET

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Title: Conformational dynamics of the ABC transporter McjD seen by single-molecule FRET
Authors: Husada, F
Bountra, K
Tasis, K
De Boer, M
Romano, M
Rebuffat, S
Beis, K
Cordes, T
Item Type: Journal Article
Abstract: ABC transporters utilize ATP for export processes to provide cellular resistance against toxins, antibiotics, and harmful metabolites in eukaryotes and prokaryotes. Based on static structure snapshots, it is believed that they use an alternating access mechanism, which couples conformational changes to ATP binding (outward‐open conformation) and hydrolysis (inward‐open) for unidirectional transport driven by ATP. Here, we analyzed the conformational states and dynamics of the antibacterial peptide exporter McjD from Escherichia coli using single‐molecule Förster resonance energy transfer (smFRET). For the first time, we established smFRET for an ABC exporter in a native‐like lipid environment and directly monitor conformational dynamics in both the transmembrane‐ (TMD) and nucleotide‐binding domains (NBD). With this, we unravel the ligand dependences that drive conformational changes in both domains. Furthermore, we observe intrinsic conformational dynamics in the absence of ATP and ligand in the NBDs. ATP binding and hydrolysis on the other hand can be observed via NBD conformational dynamics. We believe that the progress made here in combination with future studies will facilitate full understanding of ABC transport cycles.
Issue Date: 2-Nov-2018
Date of Acceptance: 5-Sep-2018
URI: http://hdl.handle.net/10044/1/64373
DOI: https://dx.doi.org/10.15252/embj.2018100056
ISSN: 0261-4189
Publisher: EMBO Press
Journal / Book Title: EMBO Journal
Volume: 37
Issue: 21
Copyright Statement: © 2018 The Authors. Published under the terms of the CC BY 4.0 license. This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Sponsor/Funder: Medical Research Council (MRC)
Funder's Grant Number: MR/N020103/1
Keywords: ABC transporter
FRET
conformational dynamics
single‐molecule studies
06 Biological Sciences
08 Information And Computing Sciences
11 Medical And Health Sciences
Developmental Biology
Publication Status: Published
Article Number: e100056
Appears in Collections:Faculty of Natural Sciences



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