Lipid Profiling Using Two-Dimensional Heteronuclear Single Quantum Coherence NMR
The use of NMR spectroscopy in lipid research has been traditionally reserved for the analysis and structural elucidation of discrete lipid molecules. Although NMR analysis of organic molecules provides a plethora of structural information that is normally unattainable by most other techniques, its use for global analysis of mixed lipid pools has been hampered by its relatively low sensitivity and overlapping of signals in the spectrum. However, the last few decades have witnessed great advancements in NMR spectroscopy that generally resulted in greater sensitivity and offered more flexibility in sampling techniques. The method discussed in this chapter describes the use of NMR for global lipidome analysis. This methodology benefits from the quantitative nature of this technique together with the abundance of the structural information it can offer, while partially overcoming the problems of low sensitivity and overlapping signals through isotope-enrichment and the use of multidimensional NMR, respectively. We have applied this method successfully to the mycobacterial lipidome as an example of an organism with a very complex and chemically diverse lipid pool. The same concept is applicable to a wide range of prokaryotes that can grow in the laboratory in well-defined growth media.
Key wordsNMR spectroscopy Mycobacteria 2D-HSQC Cell wall lipids Polyketides Glycolipids
We thank Dr Clifton Barry III, (National Institutes of Health) for providing M. tuberculosis HN-878 and HN878-▵D pks strains, Dr. Pamela L.C. Small, (University of Tennessee, Knoxville) for providing both M. marinum and M. liflindii strains, and Dr. Wei Li (University of Tennessee, Memphis) for technical assistance. We acknowledge financial support for this work from National Institutes of Health grant AI076938.
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