Identification of a broad family of lipid A late acyltransferases with non‐canonical substrate specificity
Open accessAbstract
Summary: Most Gram‐negative organisms produce lipopolysaccharide (LPS), a complex macromolecule anchored to the bacterial membrane by the lipid A moiety. Lipid A is synthesized via the R aetz pathway, a conserved nine‐step enzymatic process first characterized in Escherichia coli. The E psilonproteobacterium Helicobacter pylori uses the R aetz pathway to synthesize lipid A; however, only eight of nine enzymes in the pathway have been identified in this organism. Here, we identify the missing acyltransferase, Jhp 0255, which transfers a secondary acyl chain to the 3′‐linked primary acyl chain of lipid A, an activity similar to that of E. coli LpxM. This enzyme, reannotated as LpxJ due to limited sequence similarity with LpxM, catalyses addition of a C 12:0 or C 14:0 acyl chain to the 3′‐linked primary acyl chain of lipid A, complementing an E. coli LpxM mutant. Enzymatic assays demonstrate that LpxJ and homologues in Campylobacter jejuni and Wolinella succinogenes can act before the 2′ secondary acyltransferase, LpxL, as well as the 3‐deoxy‐d‐manno‐octulosonic acid (K do) transferase, KdtA. Ultimately, LpxJ is one member of a large class of acyltransferases found in a diverse range of organisms that lack an E. coli LpxM homologue, suggesting that LpxJ participates in lipid A biosynthesis in place of an LpxM homologue.
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Among genes involved in bacterial LPS assembly, lpxA, B, C, D, H, K and wdtA/waaA encode genes required for basic LPS assembly and tetra-acylation of lipid A, whereas lpxL facilitates LPS penta-acylation, and lpxJ and lpxM hexa-acylate LPS with differing acyl chain lengths and positions.
“Among genes involved in bacterial LPS assembly, lpxA, B, C, D, H, K and wdtA/waaA encode genes required for basic LPS assembly and tetra-acylation of lipid A, whereas lpxL facilitates LPS penta-acylation, and lpxJ and lpxM hexa-acylate LPS with differing acyl chain lengths and positions14,15.”
From Gut microbiota-derived hexa-acylated lipopolysaccharides enhance cancer immunotherapy responses (2025) · cites this paper as [15]
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Gut microbiota-derived hexa-acylated lipopolysaccharides enhance cancer immunotherapy responses
Sardar, P., Beresford-Jones, B. S., Xia, W., Shabana, O., Suyama, S., Ramos, R. J. F., Soderholm, A. T., Tourlomousis, P., Kuo, P., Evans, A. C., et al. (2025) · Nature Microbiology · doi:10.1038/s41564-025-01930-y · Cited by 49 · see citing works
- Authors
- Rubin, E. J., O'Brien, J. P., Ivanov, P. L., Brodbelt, J. S., & Trent, M. S.
- Affiliations
- The University of Texas at Austin (US)
- Corresponding author
- Trent, M. Stephen
- Venue
- Molecular Microbiology
- Publisher
- Wiley
- Published
- 2014-01-14
- Volume
- 91
- Issue
- 5
- Pages
- 887-899
- ISSN
- 0950-382X (ISSN-L)
- 1365-2958
- PMC
- PMC3947442
- OpenAlex
- W1492114039
- Cited by
- 21
- References
- 42
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2022: 1 · 2021: 2 · 2019: 2 · 2018: 3 · 2017: 3 · 2016: 4 · 2015: 1 · 2014: 1
- FWCI
- 1.33
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular Microbiology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of General Medical Sciences (R01GM103655, GM103655)
- National Institute of Allergy and Infectious Diseases (AI064184, R01AI064184)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
- Keywords
- lipid A biosynthesis, acyltransferase, Helicobacter pylori, Campylobacter jejuni, Wolinella succinogenes, lipopolysaccharide, Escherichia coli
- MeSH terms
- Acylation, Acyltransferases, Bacteria, Bacterial Proteins, Multigene Family, Genetic Complementation Test, Lipid A, Mutation, Phenotype, Substrate Specificity, Sugar Acids, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Epsilonproteobacteria
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