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Title: Crystal structure of the MOP flippase MurJ in an inward-facing conformation

Abstract

Peptidoglycan (PG) protects bacteria from osmotic lysis, and its biogenesis is a key antibiotic target. A central step in PG biosynthesis is flipping of the lipid-linked PG precursor lipid II across the cytoplasmic membrane for subsequent incorporation into PG. MurJ, part of the multidrug/oligosaccharidyl-lipid/polysaccharide (MOP) transporter superfamily, was recently shown to carry out this process. However, understanding of how MurJ flips lipid II, and of how MOP transporters operate in general, remains limited due to a lack of structural information. Here we present a crystal structure of MurJ from Thermosipho africanus in an inward-facing conformation at 2.0-Å resolution. We report a hydrophobic groove is formed by two C-terminal transmembrane helices, which leads into a large central cavity that is mostly cationic. Our studies not only provide the first structural glimpse of MurJ but also suggest that alternating access is important for MurJ function, which may be applicable to other MOP superfamily transporters.

Authors:
 [1];  [1];  [1]
  1. Duke Univ. School of Medicine, Durham, NC (United States)
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
Duke Startup Funds
OSTI Identifier:
1356413
Grant/Contract Number:  
P41GM103403; S10 RR029205
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Nature Structural & Molecular Biology
Additional Journal Information:
Journal Volume: 24; Journal Issue: 2; Journal ID: ISSN 1545-9993
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; Cellular microbiology; Membrane biophysics; Membrane proteins; X-ray crystallography

Citation Formats

Kuk, Alvin C. Y., Mashalidis, Ellene H., and Lee, Seok-Yong. Crystal structure of the MOP flippase MurJ in an inward-facing conformation. United States: N. p., 2016. Web. doi:10.1038/nsmb.3346.
Kuk, Alvin C. Y., Mashalidis, Ellene H., & Lee, Seok-Yong. Crystal structure of the MOP flippase MurJ in an inward-facing conformation. United States. https://doi.org/10.1038/nsmb.3346
Kuk, Alvin C. Y., Mashalidis, Ellene H., and Lee, Seok-Yong. 2016. "Crystal structure of the MOP flippase MurJ in an inward-facing conformation". United States. https://doi.org/10.1038/nsmb.3346. https://www.osti.gov/servlets/purl/1356413.
@article{osti_1356413,
title = {Crystal structure of the MOP flippase MurJ in an inward-facing conformation},
author = {Kuk, Alvin C. Y. and Mashalidis, Ellene H. and Lee, Seok-Yong},
abstractNote = {Peptidoglycan (PG) protects bacteria from osmotic lysis, and its biogenesis is a key antibiotic target. A central step in PG biosynthesis is flipping of the lipid-linked PG precursor lipid II across the cytoplasmic membrane for subsequent incorporation into PG. MurJ, part of the multidrug/oligosaccharidyl-lipid/polysaccharide (MOP) transporter superfamily, was recently shown to carry out this process. However, understanding of how MurJ flips lipid II, and of how MOP transporters operate in general, remains limited due to a lack of structural information. Here we present a crystal structure of MurJ from Thermosipho africanus in an inward-facing conformation at 2.0-Å resolution. We report a hydrophobic groove is formed by two C-terminal transmembrane helices, which leads into a large central cavity that is mostly cationic. Our studies not only provide the first structural glimpse of MurJ but also suggest that alternating access is important for MurJ function, which may be applicable to other MOP superfamily transporters.},
doi = {10.1038/nsmb.3346},
url = {https://www.osti.gov/biblio/1356413}, journal = {Nature Structural & Molecular Biology},
issn = {1545-9993},
number = 2,
volume = 24,
place = {United States},
year = {Mon Dec 26 00:00:00 EST 2016},
month = {Mon Dec 26 00:00:00 EST 2016}
}

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Cited by: 54 works
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Works referencing / citing this record:

A Mass‐Spectrometry‐Based Approach to Distinguish Annular and Specific Lipid Binding to Membrane Proteins
journal, January 2020


A Mass‐Spectrometry‐Based Approach to Distinguish Annular and Specific Lipid Binding to Membrane Proteins
journal, January 2020


Wzx flippases exhibiting complex O‐unit preferences require a new model for Wzx–substrate interactions
journal, May 2018


Structural basis for the coordination of cell division with the synthesis of the bacterial cell envelope
journal, September 2019


Direct observation of the influence of cardiolipin and antibiotics on lipid II binding to MurJ
journal, January 2018


Structural basis for xenobiotic extrusion by eukaryotic MATE transporter
journal, November 2017


Visualizing conformation transitions of the Lipid II flippase MurJ
journal, April 2019


A viral protein antibiotic inhibits lipid II flippase activity
journal, September 2017


Multidrug efflux pumps: structure, function and regulation
journal, July 2018


Lytic transglycosylases: concinnity in concision of the bacterial cell wall
journal, June 2017


Cell wall peptidoglycan in Mycobacterium tuberculosis: An Achilles’ heel for the TB-causing pathogen
journal, June 2019


FtsW activity and lipid II synthesis are required for recruitment of MurJ to midcell during cell division in Escherichia coli
journal, September 2018


Structural basis for xenobiotic extrusion by eukaryotic MATE transporter
journal, November 2017


Crystal structure of an intramembranal phosphatase central to bacterial cell-wall peptidoglycan biosynthesis and lipid recycling
journal, March 2018


Visualizing conformation transitions of the Lipid II flippase MurJ
journal, April 2019