文章基本信息

标题：Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony
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作者：Ping Zhou ; Vandana Chandan ; Xin Liu 等
期刊名称：JLR Papers In Press
印刷版ISSN：0022-2275
电子版ISSN：1539-7262
出版年度：2009
卷号：50
期号：9
页码：1936-1944
DOI：10.1194/jlr.D900013-JLR200
语种：English
出版社：American Society for Biochemistry and Molecular Biology
摘要：The 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.
关键词：Helicobacter pylori ; acylation ; microwave-assisted digestion ; mass spectrometry ; MALDI-TOF/TOF