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Characterization of Isomers of Lipid A from Pseudomonas aeruginosa PAO1 by Liquid Chromatography with Tandem Mass Spectrometry with Higher-Energy Collisional Dissociation and Ultraviolet Photodissociation
- Source :
- Analytical Chemistry, Analytical Chemistry, American Chemical Society, 2021, 93 (9), pp.4255-4262. ⟨10.1021/acs.analchem.0c05069⟩
- Publication Year :
- 2021
- Publisher :
- American Chemical Society (ACS), 2021.
-
Abstract
- Lipopolysaccharides (LPS) constitute the outermost layer of Gram-negative bacteria and consequently play an important role in bacterial infections. In order to address public health issues posed by Gram-negative bacteria, it is necessary to elucidate the structure of the molecular actors at the forefront of infections. LPS virulence and toxicity are partially modulated by lipid A, a hydrophobic saccharolipid that anchors LPS to the bacterial outer membrane. Understanding the lipid A structure is inherently intertwined with understanding its role as an endotoxin. Accordingly, several successful strategies incorporating tandem mass spectrometry have been applied toward the structural analysis of lipid A. Herein, a shotgun HCD strategy was applied toward the characterization of the lipid A profile of Pseudomonas aeruginosa PAO1. This analysis was enhanced by the development of an LC-MS/MS approach to eliminate isomeric signals in the MS/MS spectra that confounded characterization. Importantly, combining reverse phase chromatography with HCD and ultraviolet photodissociation analyses of the lipid A profile revealed the presence of previously unreported lipid A acyl chain positional isomers. Altogether, these strategies provide the most in-depth structural and molecular characterization of PAO1 lipid A to date.
- Subjects :
- Ions
Mass spectrometry
biology
Chemistry
Acyls
010401 analytical chemistry
Photodissociation
[CHIM.MATE]Chemical Sciences/Material chemistry
Reversed-phase chromatography
010402 general chemistry
biology.organism_classification
Tandem mass spectrometry
Lipids
01 natural sciences
0104 chemical sciences
Analytical Chemistry
Lipid A
Chemical structure
Biochemistry
Structural isomer
lipids (amino acids, peptides, and proteins)
Saccharolipid
Bacterial outer membrane
Bacteria
Subjects
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 93
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....1854d72dec11d535c52d5b98fad97b04