Document Detail

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.
MedLine Citation:
PMID:  15371346     Owner:  NLM     Status:  MEDLINE    
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 DeltaembB 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.
Robin E B Lee; Wei Li; Delphi Chatterjee; Richard E Lee
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S.     Date:  2004-09-15
Journal Detail:
Title:  Glycobiology     Volume:  15     ISSN:  0959-6658     ISO Abbreviation:  Glycobiology     Publication Date:  2005 Feb 
Date Detail:
Created Date:  2004-12-30     Completed Date:  2005-06-16     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9104124     Medline TA:  Glycobiology     Country:  England    
Other Details:
Languages:  eng     Pagination:  139-51     Citation Subset:  IM    
Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, 847 Monroe Ave. Rm. 327, Memphis, TN 38163, USA.
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MeSH Terms
Carbohydrate Sequence / drug effects,  genetics
Cell Wall / chemistry,  genetics
Ethambutol / chemistry*
Galactans / chemistry*,  genetics
Gene Deletion*
Molecular Sequence Data
Mycobacterium bovis / chemistry*,  genetics
Mycobacterium smegmatis / chemistry*,  genetics
Nuclear Magnetic Resonance, Biomolecular
Polysaccharides / chemistry*,  genetics
Polysaccharides, Bacterial / chemistry*,  genetics
Grant Support
Reg. No./Substance:
0/Galactans; 0/Lipopolysaccharides; 0/Polysaccharides; 0/Polysaccharides, Bacterial; 0/lipoarabinomannan; 11078-27-6/araban; 74-55-5/Ethambutol; 9036-66-2/arabinogalactan

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