Comprehensive analysis of exported proteins from Mycobacterium tuberculosis H37Rv

• Published in: Proteomics (Weinheim) Vol. 7; no. 10; pp. 1702 - 1718
• Main Authors: Målen, Hiwa, Berven, Frode S., Fladmark, Kari E., Wiker, Harald G.
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.05.2007; WILEY‐VCH Verlag
• Subjects: Amino Acid Sequence; Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; Electrophoresis, Gel, Two-Dimensional; Liquid chromatography-tandem mass spectrometry; Mass spectra; Molecular Sequence Data; Molecular Weight; Mycobacterium tuberculosis; Mycobacterium tuberculosis - chemistry; Secreted protein; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Two-dimensional gel electrophoresis
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.200600853

Proteins secreted by Mycobacterium tuberculosis play an essential role in the pathogenesis of tuberculosis. The culture filtrates of M. tuberculosis H37Rv made by Sadamu Nagai (Japan), are considerably enriched for secreted proteins compared to other culture filtrates. Complementary approaches were used to identify the secreted proteins in these culture filtrates: (i) 2‐DE combined with MALDI‐TOF MS and (ii) LC coupled MS/MS. Peptides derived from a total of 257 proteins were identified of which 144 were identified by more than one peptide. Several members of the immunologically important early secretory antigenic target‐6 (ESAT‐6) family of proteins were found to be major components. The majority of the identified proteins, 159 (62%), were predicted to be exported through the general secretory pathway. We experimentally verified that the signal peptides, which mediate translocation through the cell membrane, had been removed in 41 of the identified proteins, and in 35 of those, there was an AXA motif N‐terminally to the cleavage site, showing that this motif is important for the recognition and cleavage of signal peptides in mycobacteria. A large fraction of the secreted proteins were unknown, suggesting that we have mapped an unexplored part of the exported proteome of M. tuberculosis.∁