A rapid approach to lipid profiling of mycobacteria using 2D HSQC NMR maps

Engy A. Mahrous, Robin B. Lee, and Richard E. Lee

Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163

Corresponding Author: relee{at}utmem.edu

Mycobacteria, including M. tuberculosis, are characterized by a unique cell wall rich in complex lipids, glycolipids, polyketides and terpenoids. Many of these metabolites have been shown to play important roles in mycobacterial virulence and their inherent resistance to many antibiotics. Herein, we report the development of a new simple method for global analysis of these metabolites utilizing two-dimensional 1H-13C heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance. The major advantages of this method are: the small amount of sample and the minimal sample manipulation required; a relatively short procedural time; the ability to rapidly attain a qualitative and quantitative lipid profile of a mycobacterial sample in which the majority of the clinically relevant lipids can be observed simultaneously. The effectiveness of this method is demonstrated in four different areas of major concern to the mycobacterial research community (i) Adaptive changes in the cell wall lipids as a result of drug treatment (ii) Analysis of gene function (iii) Characterization of new mycobacterial species (iv) Analysis of the production of virulence factors in clinical isolates of M. tuberculosis. This method is complementary to mass spectrometry based lipidomic technologies and provides an urgently needed tool to gain a better understanding the role of lipids in mycobacteria pathogenesis.

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