Definition of novel cell envelope associated proteins in Triton X-114 extracts of Mycobacterium tuberculosis H37Rv.
BACKGROUND
Membrane- and membrane-associated proteins are important for the pathogenicity of bacteria. We have analysed the content of these proteins in virulent Mycobacterium tuberculosis H37Rv using Triton X-114 detergent-phase separation for extraction of lipophilic proteins, followed by their identification with high resolution mass spectrometry.


RESULTS
In total, 1417 different proteins were identified. In silico analysis of the identified proteins revealed that 248 proteins had at least one predicted trans-membrane region. Also, 64 of the identified proteins were predicted lipoproteins, and 54 proteins were predicted as outer membrane proteins. Three-hundred-and-ninety-five of the observed proteins, including 91 integral membrane proteins were described for the first time. Comparison of abundance levels of the identified proteins was performed using the exponentially modified protein abundance index (emPAI) which takes into account the number of the observable peptides to the number of experimentally observed peptide ions for a given protein. The outcome showed that among the membrane-and membrane-associated proteins several proteins are present with high relative abundance. Further, a close examination of the lipoprotein LpqG (Rv3623) which is only detected in the membrane fractions of M. tuberculosis but not in M. bovis, revealed that the homologous gene in M. bovis lack the signal peptide and lipobox motif, suggesting impaired export to the membrane.


CONCLUSIONS
Altogether, we have identified a substantial proportion of membrane- and membrane-associated proteins of M. tuberculosis H37Rv, compared the relative abundance of the identified proteins and also revealed subtle differences between the different members of the M. tuberculosis complex.
Målen
Hiwa
H
Section for Microbiology and Immunology, the Gade Institute, University of Bergen, Bergen, Norway.
Pathak
Sharad
S
Søfteland
Tina
T
de Souza
Gustavo A
GA
Wiker
Harald G
HG
D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification*; D002462:Cell Membrane / Q000737:chemistry* / Q000187:drug effects; D019295:Computational Biology; D003902:Detergents / Q000494:pharmacology*; D008074:Lipoproteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D013058:Mass Spectrometry; D009169:Mycobacterium tuberculosis / Q000737:chemistry* / Q000187:drug effects; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology*; D021382:Protein Sorting Signals / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D003902:Detergents; D008074:Lipoproteins; D021382:Protein Sorting Signals; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40