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Lookup NU author(s): Dr Kesha Josts
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
The ATP-binding cassette transporter MsbA is a lipid flippase, translocating lipid A, glycolipids, and lipopolysaccharides from the inner to the outer leaflet of the inner membrane of Gram-negative bacteria. It has been used as a model system for time-resolved structural studies as several MsbA structures in different states and reconstitution systems (detergent/nanodiscs/peptidiscs) are available. However, due to the limited resolution of the available structures, detailed structural information on the bound nucleotides has remained elusive. Here, we have reconstituted MsbA in saposin A–lipoprotein nanoparticles (Salipro) and determined the structure of ADP-vanadate-bound MsbA by single-particle cryo-electron microscopy to 3.5 Å resolution. This procedure has resulted in significantly improved resolution and enabled us to model all side chains and visualise detailed ADP-vanadate interactions in the nucleotide-binding domains. The approach may be applicable to other dynamic membrane proteins.
Author(s): Kehlenbeck DM, Traore DA, Josts I, Sander S, Moulin M, Haertlein M, Prevost S, Forsyth VT, Tidow H
Publication type: Article
Publication status: Published
Journal: The FEBS journal
Year: 2022
Volume: 289
Issue: 10
Pages: 2959-2970
Print publication date: 17/05/2022
Online publication date: 17/12/2021
Acceptance date: 16/12/2021
Date deposited: 12/06/2024
ISSN (print): 1742-464X
ISSN (electronic): 1742-4658
Publisher: Wiley-Blackwell Publishing Ltd.
URL: https://doi.org/10.1111/febs.16327
DOI: 10.1111/febs.16327
Data Access Statement: The cryo-EM map and the atomic coordinates are available in the Electron Microscopy Data Bank and the Protein Data Bank under accession number EMDB-12145 and 7BCW, respectively. All other relevant data generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
PubMed id: 34921499
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