Glycobiology Advance Access originally published online on September 15, 2004
Glycobiology 2005 15(2):139-151; doi:10.1093/glycob/cwh150
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Glycobiology vol. 15 no. 2 © Oxford University Press 2005; all rights reserved.
Rapid structural characterization of the arabinogalactan and lipoarabinomannan in live mycobacterial cells using 2D and 3D HR-MAS NMR: structural changes in the arabinan due to ethambutol treatment and gene mutation are observed
2 Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, 847 Monroe Ave. Rm. 327, Memphis, TN 38163; 3 Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523
1 To whom correspondence should be addressed; e-mail: relee{at}utmem.edu
Received on July 21, 2004; revised on September 13, 2004; accepted on September 14, 2004
Mycobacteria possess a unique, highly evolved, carbohydrate- and lipid-rich cell wall that is believed to be important for their survival in hostile environments. Until now, our understanding of mycobacterial cell wall structure has been based upon destructive isolation and fragmentation of individual cell wall components. This study describes the observation of the major cell wall structures in live, intact mycobacteria using 2D and 3D high-resolution magic-angle spinning (HR-MAS) nuclear magnetic resonance (NMR). As little as 20 mg (wet weight) of [13C]-enriched cells were required to produce a whole-cell spectra in which discrete cross-peaks corresponding to specific cell wall components could be identified. The most abundant signals of the arabinogalactan (AG) and lipoarabinomannan (LAM) were assigned in the HR-MAS NMR spectra by comparing the 2D and 3D NMR whole-cell spectra with the spectra of purified cellular components. This study confirmed that the structures of the AG and LAM moieties in the cell wall of live mycobacteria are consistent with structural reports in the literature, which were obtained via degradative analysis. Most important, by using intact cells it was possible to directly demonstrate the effects of ethambutol on the mycobacterial cell wall polysaccharides, characterize the effects of embB gene knockout in the M. smegmatis 
embB mutant, and observe differences in the cell wall structures of two mycobacterial species (M. bovis BCG and M. smegmatis.) Herein, we show that HR-MAS NMR is a powerful, rapid, nondestructive technique to monitor changes in the complex, carbohydrate-rich cell wall of live mycobacterial cells.
Key words: arabinogalactan / HCCH-TOCSY / HR-MAS NMR / lipoarabinomannan / mycobacteria
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  H. Safi, B. Sayers, M. H. Hazbon, and D. Alland Transfer of embB Codon 306 Mutations into Clinical Mycobacterium tuberculosis Strains Alters Susceptibility to Ethambutol, Isoniazid, and Rifampin Antimicrob. Agents Chemother., June 1, 2008; 52(6): 2027 - 2034. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. A. Mahrous, R. B. Lee, and R. E. Lee A rapid approach to lipid profiling of mycobacteria using 2D HSQC NMR maps J. Lipid Res., February 1, 2008; 49(2): 455 - 463. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Shi, S. Berg, A. Lee, J. S. Spencer, J. Zhang, V. Vissa, M. R. McNeil, K.-H. Khoo, and D. Chatterjee The Carboxy Terminus of EmbC from Mycobacterium smegmatis Mediates Chain Length Extension of the Arabinan in Lipoarabinomannan J. Biol. Chem., July 14, 2006; 281(28): 19512 - 19526. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Dong, S. Bhamidi, M. Scherman, Y. Xin, and M. R. McNeil Development of a Quantitative Assay for Mycobacterial Endogenous Arabinase and Ensuing Studies of Arabinase Levels and Arabinan Metabolism in Mycobacterium smegmatis Appl. Envir. Microbiol., April 1, 2006; 72(4): 2601 - 2605. [Abstract] [Full Text] [PDF]
|
|