Proteomic definition of the cell wall of Mycobacterium tuberculosis.
The cell envelope of Mycobacterium tuberculosis (Mtb) is complex and diverse; composed of proteins intermingled in a matrix of peptidoglycan, mycolic acids, lipids, and carbohydrates. Proteomic studies of the Mtb cell wall have been limited; nonetheless, the characterization of resident and secreted proteins associated with the cell wall are critical to understanding bacterial survival and immune modulation in the host. In this study, the cell wall proteome was defined in order to better understand its unique biosynthetic and secretion processes. Mtb cell wall was subjected to extraction with organic solvents to remove noncovalently bound lipids and lipoglycans and remaining proteins were solubilized with either SDS, Guanidine-HCl, or TX-114. These extracts were analyzed by two-dimensional gel electrophoresis and mass-spectrometry and resulted in the identification of 234 total proteins. The lipoproteome of Mtb, enriched in the TX-114 extract, was further resolved by multidimensional chromatography and mass spectrometry to identify an additional 294 proteins. A query of the 528 total protein identifications against Neural Network or Hidden Markov model algorithms predicted secretion signals in 87 proteins. Classification of these 528 proteins also demonstrated that 35% are involved in small molecule metabolism and 25% are involved in macromolecule synthesis and degradation building upon evidence that the Mtb cell wall is actively engaged in mycobacterial survival and remodeling.
Wolfe
Lisa M
LM
Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, USA.
Mahaffey
Spencer B
SB
Kruh
Nicole A
NA
Dobos
Karen M
KM
D001426:Bacterial Proteins / Q000032:analysis*; D002473:Cell Wall / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry; D009169:Mycobacterium tuberculosis / Q000737:chemistry* / Q000648:ultrastructure; D040901:Proteomics / Q000379:methods*; D012995:Solubility; D012997:Solvents
D001426:Bacterial Proteins; D012997:Solvents
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