Mapping the architecture of the initiating phosphoglycosyl transferase from S. enterica O-antigen biosynthesis in a liponanoparticle
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Abstract
Bacterial cell surface glycoconjugates are critical for cell survival and for interactions between bacteria and their hosts. Consequently, the pathways responsible for their biosynthesis have untapped potential as therapeutic targets. The localization of many glycoconjugate biosynthesis enzymes to the membrane represents a significant challenge for expressing, purifying, and characterizing these enzymes. Here, we leverage cutting-edge detergent-free methods to stabilize, purify, and structurally characterize WbaP, a phosphoglycosyl transferase (PGT) from the Salmonella enterica (LT2) O-antigen biosynthesis. From a functional perspective, these studies establish WbaP as a homodimer, reveal the structural elements responsible for dimerization, shed light on the regulatory role of a domain of unknown function embedded within WbaP, and identify conserved structural motifs between PGTs and functionally unrelated UDP-sugar dehydratases. From a technological perspective, the strategy developed here is generalizable and provides a toolkit for studying other classes of small membrane proteins embedded in liponanoparticles beyond PGTs.
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Fast and accurate protein structure search with Foldseek
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- Authors
- Dodge, G. J., Anderson, A. J., He, Y., Liu, W., Viner, R., & Imperiali, B.
- Affiliations
- University of Alberta (CA)
- Corresponding author
- Imperiali, Barbara
- Venue
- eLife
- Publisher
- eLife Sciences Publications, Ltd
- Published
- 2024-02-15
- Volume
- 12
- ISSN
- 2050-084X (ISSN-L)
- PMC
- PMC10942596
- OpenAlex
- W4388971678
- Cited by
- 9
- References
- 6
- Citations by year
- 2026: 3 · 2025: 4 · 2024: 2
- FWCI
- 0.92
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- eLife (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Institutes of Health (GM-039334, GM-131627, GM-134576)
- Topics
- Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- Salmonella enterica, O-antigen biosynthesis, homodimerization, domain of unknown function
- MeSH terms
- Cell Membrane, Transferases, O Antigens, Salmonella enterica, Carbohydrate Metabolism
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