Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony[S]
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 n...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2009-09-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S002222752030746X |
| _version_ | 1818601064608301056 |
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| author | Ping Zhou Vandana Chandan Xin Liu Kenneth Chan Eleonora Altman Jianjun Li |
| author_facet | Ping Zhou Vandana Chandan Xin Liu Kenneth Chan Eleonora Altman Jianjun Li |
| author_sort | Ping Zhou |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-16T12:45:27Z |
| format | Article |
| id | doaj.art-5145ad443ddc4dc6b2a3d5c88c3e8f0b |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-16T12:45:27Z |
| publishDate | 2009-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-5145ad443ddc4dc6b2a3d5c88c3e8f0b2022-12-21T22:31:19ZengElsevierJournal of Lipid Research0022-22752009-09-0150919361944Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony[S]Ping Zhou0Vandana Chandan1Xin Liu2Kenneth Chan3Eleonora Altman4Jianjun Li5Institute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, Canada; College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, Peoples' Republic of ChinaInstitute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, CanadaInstitute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, CanadaInstitute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, CanadaInstitute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, CanadaTo whom correspondence should be addressed.; Institute for Biological Sciences, National Research Council Canada, Ottawa, Ontario, CanadaThe 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.http://www.sciencedirect.com/science/article/pii/S002222752030746XHelicobacter pyloriacylationmicrowave-assisted digestionmass spectrometryMALDI-TOF/TOF |
| spellingShingle | Ping Zhou Vandana Chandan Xin Liu Kenneth Chan Eleonora Altman Jianjun Li Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony[S] Journal of Lipid Research Helicobacter pylori acylation microwave-assisted digestion mass spectrometry MALDI-TOF/TOF |
| title | Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony[S] |
| title_full | Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony[S] |
| title_fullStr | Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony[S] |
| title_full_unstemmed | Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony[S] |
| title_short | Microwave-assisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony[S] |
| title_sort | microwave assisted sample preparation for rapid and sensitive analysis of h pylori lipid a applicable to a single colony s |
| topic | Helicobacter pylori acylation microwave-assisted digestion mass spectrometry MALDI-TOF/TOF |
| url | http://www.sciencedirect.com/science/article/pii/S002222752030746X |
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