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Mycobacteria, including Mycobacterium 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.

Here, we report the development of a new simple method for global analysis of these metabolites using two-dimensional ¹H-¹³C heteronuclear single quantum coherence nuclear magnetic resonance.

The major advantages of this method are as follows: the small amount of sample and the minimal sample manipulation required; a relatively short procedural time; and 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 cell wall lipids as a result of drug treatment; i
• i) analysis of gene function;
• iii) characterization of new mycobacterial species; and
• 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 of the role of lipids in mycobacteria pathogenesis.

Supplementary key words two-dimensional ¹H-¹³C heteronuclear single quantum coherence nuclear magnetic resonance • lipidomics • phenolic glycolipids • tuberculosis

Engy A. Mahrous, Robin B. Lee and Richard E. Lee¹

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

To whom correspondence should be addressed. e-mail: relee@utmem.edu