出版社：American Society for Biochemistry and Molecular Biology
摘要：Here, we present an improved method for sensitive profilingof lipids in a single high-performance liquid chromatography-electrosprayionization-quadrupole time of flight mass spectrometry experiment.The approach consists of i) sensitive isocratic elution, whichtakes advantage of C18 column material that is resistant toincreased pH values induced by piperidine, ii) chemometric alignmentof mass spectra followed by differential analysis of ion intensities,and iii) semiquantitative analysis of extracted ion chromatogramsof interest. A key advantage of this method is its wide applicabilityto extracts that harbor lipids of considerable chemical complexity.The method allows qualitative and semiquantitative analysisof fatty acyls, glycerophospholipids (such as glycerophosphatidylinositols,glycerophosphatidylserines, and glycerophosphatidylcholinesin brain extracts), phosphatidylinositol mannosides, acylatedglycerophospholipids, sphingolipids (including ceramides andgangliosides in brain extracts), and, for the first time withESI, prenols and mycolic acids (MAs). MAs are targets in antimycobacterialtherapy, and they play an important immunomodulatory role duringhost-pathogen interactions. We compared high-resolution massspectra of MAs derived from Mycobacterium bovis Bacille Camette-Guérinduring entry into nonreplicative conditions induced by oxygendeprivation (hypoxic dormancy). Although the overall compositionis not drastically altered, there are pronounced differencesin individual MAs. -MAs accumulate during entry into dormancy,whereas a subpopulation of keto-MAs is almost entirely eliminated.This effect is reversed upon resuscitation of dormant mycobacteria.These results provide detailed chemical information with relevanceto drug development and immunobiology of mycobacteria.Supplementary key words lipidomics • liquid chromatography • mass spectrometry • mycobacteria • mycolic acids • drug development • host-pathogen interactions • hypoxic dormancy
Abbreviations: BCG, Bacille Camette-Guérin; COW, correlation-optimized warping; MA, mycolic acid; PIP, phosphatidylinositol polyphosphate; Q-TOF, quadrupole/time of flight
-MAs accumulate during entry into dormancy, whereas a subpopulation of keto-MAs is almost entirely eliminated. This effect is reversed upon resuscitation of dormant mycobacteria. These results provide detailed chemical information with relevance to drug development and immunobiology of mycobacteria.
Supplementary key words lipidomics • liquid chromatography • mass spectrometry • mycobacteria • mycolic acids • drug development • host-pathogen interactions • hypoxic dormancy
Abbreviations: BCG, Bacille Camette-Guérin; COW, correlation-optimized warping; MA, mycolic acid; PIP, phosphatidylinositol polyphosphate; Q-TOF, quadrupole/time of flight
