Structure of WzxE the lipid III flippase for Enterobacterial Common Antigen polysaccharide
The enterobacterial common antigen (ECA) is conserved in Gram-negative bacteria of the <i>Enterobacterales</i> order although its function is debated. ECA biogenesis depends on the Wzx/Wzy-dependent strategy whereby the newly synthesized lipid-linked repeat units, lipid III, are transferred across the inner membrane by the lipid III flippase WzxE. WzxE is part of the Wzx family and required in many glycan assembly systems, but an understanding of its molecular mechanism is hindered due to a lack of structural evidence. Here, we present the first X-ray structures of WzxE from <i>Escherichia coli</i> in complex with nanobodies. Both inward- and outward-facing conformations highlight two pairs of arginine residues that move in a reciprocal fashion, enabling flipping. One of the arginine pairs coordinated to a glutamate residue is essential for activity along with the C-terminal arginine rich tail located close to the entrance of the lumen. This work helps understand the translocation mechanism of the Wzx flippase family.
Authors
- Jan Steyaert (ORCID: https://orcid.org/0000-0002-3825-874X)
- Els Pardon (ORCID: https://orcid.org/0000-0002-2466-0172)
- Peter J. Harrison (ORCID: https://orcid.org/0000-0001-5167-0547)
- Neil G. Paterson (ORCID: https://orcid.org/0000-0003-4292-6971)
- Miriam Weckener (ORCID: https://orcid.org/0000-0002-1419-9378)
- Kamel El Omari (ORCID: https://orcid.org/0000-0003-3506-6045)
- Andrew M. Giltrap (ORCID: https://orcid.org/0000-0001-8960-553X)
- Micah Lee (ORCID: https://orcid.org/0000-0001-8447-0314)
- Benjamin G. Davis (ORCID: https://orcid.org/0000-0002-5056-407X)
- S.A. McMahon (ORCID: https://orcid.org/0000-0003-4273-8364)
- Huanting Liu (ORCID: https://orcid.org/0000-0001-5953-9367)
- Chris Whitfield (ORCID: https://orcid.org/0000-0002-5267-7027)
- Tanisha Teelucksingh (ORCID: https://orcid.org/0000-0002-1151-8433)
- James H. Naismith (ORCID: https://orcid.org/0000-0001-6744-5061)
- Armin Wagner (ORCID: https://orcid.org/0000-0001-8995-7324)
- Audrey Le Bas (ORCID: https://orcid.org/0000-0002-1100-6833)
- Philip N. Ward (ORCID: https://orcid.org/0000-0003-2546-3426)
- Andrew Quigley (ORCID: https://orcid.org/0000-0002-5022-9845)
- Bradley R. Clarke (ORCID: https://orcid.org/0000-0003-2233-1268)
- John H. Beale (ORCID: https://orcid.org/0000-0003-1171-2428)
- Ramona Duman (ORCID: https://orcid.org/0000-0002-3566-8698)
- Hui Liu (ORCID: https://orcid.org/0009-0009-4988-9561)
- Vitaliy Mykhaylyk
Institutions
- Vrije Universiteit Brussel (BE)
- University of St Andrews (GB)
- Theodore Roosevelt High School (US)
- Diamond Light Source (GB)
- University of Oxford (GB)
- Research Complex at Harwell (GB)
- Rosalind Franklin Institute (GB)
- Université Bourgogne Franche-Comté (FR)
- VIB-VUB Center for Structural Biology (BE)
- University of Guelph (CA)
Publication Details
- Journal
- Open Biology
- Published
- 2025-01-01
- DOI
- https://doi.org/10.1098/rsob.240310
- Citations
- 8
- Primary Topic
- Escherichia coli research studies
- Type
- article
- Field-Weighted Citation Impact
- 14.38
Funders
- Wellcome Trust
- Diamond Light Source
- Canada Research Chairs
- Canadian Institutes of Health Research
- Engineering and Physical Sciences Research Council