Microwave-assisted sample preparation for rapid and sensitive analysis of Helicobacter pylori lipid A applicable to a single colony

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DOIResolve DOI: http://doi.org/10.1194/jlr.D900013-JLR200
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TypeArticle
Journal titleJournal of Lipid Research
Volume50
Issue9
Pages19361944; # of pages: 9
SubjectHelicobacter pylori; acylation; microwave-assisted digestion; mass spectrometry; MALDI-TOF/TOF
AbstractThe lipid A of Gram-negative bacteria plays a major role in the pathogenesis of bacterial infections. Lipid A diversity is observed both in the number and length of fatty acid side chains and in the presence of terminal phosphate residues and associated modifications. In this report, we describe a new sample preparation method based on microwave-assisted enzymatic digestion and detergent-free mild hydrolysis, in conjunction with a MALDI-time-of-flight (TOF)/TOF analysis, to determine the structures of lipid A from Helicobacter pylori . The total time for sample preparation and mass spectrometric analysis is within 2 h and applicable to profiling the lipid A structures from dried bacterial cells on as little as 1 µg. The reliability of the technique was further demonstrated through the analysis of the lipid A from bacterial cells of different H. pylori strains. The phosphorylation and acylation patterns of lipid A could be elucidated using material from a single colony. Furthermore, we found unusual heptaacyl lipid A species present in H. pylori mutant that have not been previously reported, although the abundance was relatively low. The present study provides the first characterization of the lipid A component from a single bacterial colony sample by mass spectrometry.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); NRC Institute for Biological Sciences
Peer reviewedYes
NPARC number15329308
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Record identifier625f2850-6018-45c8-b3b8-1409129ce650
Record created2010-05-21
Record modified2016-05-09
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