Summary
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Goren, M. B., O. Brokl, and W. B. Schaefer. 1974. Lipids of putative relevance to virulence in Mycobacterium tuberculosis: phthiocerol dimycocerosate and the attenuation indicator lipid. Infection and immunity 9:150–158.
Goren, M. B., O. Brokl, and W. B. Schaefer. 1974. Lipids of putative relevance to virulence in Mycobacterium tuberculosis: correlation of virulence with elaboration of sulfatides and strongly acidic lipids. Infection and immunity 9:142–149.
Kato, M., M. Kusunose, K. Miki, K. Matsunaga, and Y. Yamamura. 1959. The mechanism of the toxicity of cord factor. The American review of respiratory disease 80:240–248.
Etemadi, A. H. 1967. [Mycolic acids. Structure, biogenesis and phylogenetic value]. Exposes annuels de biochimie medicale 28:77–109.
Etemadi, A. H. 1967. [Structural and biogenetic correlations of mycolic acids in relation to the phylogenesis of various genera of Actinomycetales]. Bulletin de la Societe de chimie biologique 49:695–706.
Lechevalier, M. P., A. C. Horan, and H. Lechevalier. 1971. Lipid composition in the classification of nocardiae and mycobacteria. Journal of bacteriology 105:313–318.
Minnikin, D. E., L. Alshamaony, and M. Goodfellow. 1975. Differentiation of Mycobacterium, Nocardia, and related taxa by thin-layer chromatographic analysis of whole-organism methanolysates. Journal of general microbiology 88:200–204.
Dobson, G., D. E. Minnikin, S. M. Minnikin, M. Parlett, M. Goodfellow, M. Ridell, and M. Magnusson. 1985. Systematic analysis of complex mycobacterial lipids. In Chemical methods in bacterial systematics. M. Goodfellow, D. E. Minnikin(eds). Academic, London. 237–2654.
Jain, M., C. J. Petzold, M. W. Schelle, M. D. Leavell, J. D. Mougous, C. R. Bertozzi, J. A. Leary, and J. S. Cox. 2007. Lipidomics reveals control of Mycobacterium tuberculosis virulence lipids via metabolic coupling. Proceedings of the National Academy of Sciences of the United States of America 104:5133–5138.
Shui, G., A. K. Bendt, K. Pethe, T. Dick, and M. R. Wenk. 2007. Sensitive profiling of chemically diverse bioactive lipids. Journal of lipid research 48:1976–1984.
Mahrous, E. A., R. B. Lee, and R. E. Lee. 2008. A rapid approach to lipid profiling of mycobacteria using 2D HSQC NMR maps. Journal of lipid research 49:455–463.
WHO report 2008 Global tuberculosis control surveillance, planning, financing. WHO. 17–20.
Brennan, P. J. and H. Nikaido. 1995. The envelope of mycobacteria. Annual review of biochemistry 64:29–63.
Puzo, G. 1990. The carbohydrate- and lipid-containing cell wall of mycobacteria, phenolic glycolipids: structure and immunological properties. Critical reviews in microbiology 17:305–327.
Karakousis, P. C., W. R. Bishai, and S. E. Dorman. 2004. Mycobacterium tuberculosis cell envelope lipids and the host immune response. Cellular microbiology 6:105–116.
Ortalo-Magne, A., A. Lemassu, M. A. Laneelle, F. Bardou, G. Silve, P. Gounon, G. Marchal, and M. Daffe. 1996. Identification of the surface-exposed lipids on the cell envelopes of Mycobacterium tuberculosis and other mycobacterial species. Journal of bacteriology 178:456–461.
Villeneuve, C., G. Etienne, V. Abadie, H. Montrozier, C. Bordier, F. Laval, M. Daffe, I. Maridonneau-Parini, and C. Astarie-Dequeker. 2003. Surface-exposed glycopeptidolipids of Mycobacterium smegmatis specifically inhibit the phagocytosis of mycobacteria by human macrophages. Identification of a novel family of glycopeptidolipids. The Journal of biological chemistry 278:51291–51300.
Villeneuve, C., M. Gilleron, I. Maridonneau-Parini, M. Daffe, C. Astarie-Dequeker, and G. Etienne. 2005. Mycobacteria use their surface-exposed glycolipids to infect human macrophages through a receptor-dependent process. Journal of lipid research 46:475–483.
Acknowledgments
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Mahrous, E.A., Lee, R.B., Lee, R.E. (2009). Lipid Profiling Using Two-Dimensional Heteronuclear Single Quantum Coherence NMR. In: Armstrong, D. (eds) Lipidomics. Methods in Molecular Biology, vol 579. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-322-0_5
Download citation
DOI: https://doi.org/10.1007/978-1-60761-322-0_5
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-321-3
Online ISBN: 978-1-60761-322-0
eBook Packages: Springer Protocols