Discovery of Sulfated Metabolites in Mycobacteria with a Genetic and Mass Spectrometric Approach

The study of the metabolome presents numerous challenges, first among them being the cataloging of its constituents. A step in this direction will be the development of tools to identify metabolites that share common structural features. The importance of sulfated molecules in cell-cell communicatio...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2002-12, Vol.99 (26), p.17037-17042
Main Authors: Mougous, Joseph D., Leavell, Michael D., Senaratne, Ryan H., Leigh, Clifton D., Williams, Spencer J., Riley, Lee W., Leary, Julie A., Bertozzi, Carolyn R.
Format: Article
Language:English
Subjects:
Average linear density
Bacteria
Biological Sciences
Cellular metabolism
Cysteine
Disaccharides - metabolism
Genetic Engineering
Genetics
Glycopeptides
Inositol
Ions
Mass Spectrometry
Metabolism
Metabolites
Microbiology
Molecules
Mycobacterium smegmatis - metabolism
Mycobacterium tuberculosis
Mycobacterium tuberculosis - metabolism
Pyrazoles - metabolism
Scientific imaging
Sulfates
Sulfates - metabolism
Sulfation
Sulfhydryl Compounds - metabolism
Sulfoglycosphingolipids - metabolism
Sulfur
Sulfur Isotopes
Tuberculosis
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Summary:The study of the metabolome presents numerous challenges, first among them being the cataloging of its constituents. A step in this direction will be the development of tools to identify metabolites that share common structural features. The importance of sulfated molecules in cell-cell communication motivated us to develop a rapid two-step method for identifying these metabolites in microorganisms, particularly in pathogenic mycobacteria. Sulfur-containing molecules were initially identified by mass spectral analysis of cell extracts from bacteria labeled metabolically with a stable sulfur isotope (34SO4 2-). To differentiate sulfated from reduced-sulfur-containing molecules, we employed a mutant lacking the reductive branch of the sulfate assimilation pathway. In these sulfur auxotrophs, heavy sulfate is channeled exclusively into sulfated metabolites. The method was applied to the discovery of several new sulfated molecules in Mycobacterium tuberculosis and Mycobacterium smegmatis. Because a sulfur auxotrophic strain is the only requirement of the approach, many microorganisms can be studied in this manner. Such genetic engineering in combination with stable isotopic labeling can be applied to various metabolic pathways and their products.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.252514899