UVliPiD: A UVPD-Based Hierarchical Approach for De Novo Characterization of Lipid A Structures

The lipid A domain of the endotoxic lipopolysaccharide layer of Gram-negative bacteria is comprised of a diglucosamine backbone to which a variable number of variable length fatty acyl chains are anchored. Traditional characterization of these tails and their linkages by nuclear magnetic resonance (...

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Published inAnalytical chemistry (Washington) Vol. 88; no. 3; pp. 1812 - 1820
Main Authors Morrison, Lindsay J, Parker, W. Ryan, Holden, Dustin D, Henderson, Jeremy C, Boll, Joseph M, Trent, M. Stephen, Brodbelt, Jennifer S
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 02.02.2016
Subjects
Automation
Bacteria
Carbohydrates
E coli
Escherichia coli - chemistry
Fatty acids
Gram-negative bacteria
Lipid A - chemistry
Lipids
Mass Spectrometry
Nuclear magnetic resonance
Protein Conformation
Spectra
Strategy
Ultraviolet Rays
Utilities
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Summary:The lipid A domain of the endotoxic lipopolysaccharide layer of Gram-negative bacteria is comprised of a diglucosamine backbone to which a variable number of variable length fatty acyl chains are anchored. Traditional characterization of these tails and their linkages by nuclear magnetic resonance (NMR) or mass spectrometry is time-consuming and necessitates databases of pre-existing structures for structural assignment. Here, we introduce an automated de novo approach for characterization of lipid A structures that is completely database-independent. A hierarchical decision-tree MS n method is used in conjunction with a hybrid activation technique, UVPDCID, to acquire characteristic fragmentation patterns of lipid A variants from a number of Gram-negative bacteria. Structural assignments are derived from integration of key features from three to five spectra and automated interpretation is achieved in minutes without the need for pre-existing information or candidate structures. The utility of this strategy is demonstrated for a mixture of lipid A structures from an enzymatically modified E. coli lipid A variant. A total of 27 lipid A structures were discovered, many of which were isomeric, showcasing the need for a rapid de novo approach to lipid A characterization.
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ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.5b04098