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Immunogenic membrane-associated proteins of Mycobacterium tuberculosis revealed by proteomics
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| Zeitschriftentitel: | Microbiology |
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| Personen und Körperschaften: | , , , , , , , |
| In: | Microbiology, 151, 2005, 7, S. 2411-2419 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Microbiology Society
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| Schlagwörter: |
| Zusammenfassung: | <jats:p>Membrane-associated proteins of<jats:italic>Mycobacterium tuberculosis</jats:italic>offer a challenge, as well as an opportunity, in the quest for better therapeutic and prophylactic interventions against tuberculosis. The authors have previously reported that extraction with the detergent Triton X-114 (TX-114) is a useful step in proteomic analysis of mycobacterial cell membranes, and detergent-soluble membrane proteins of mycobacteria are potent stimulators of human T cells. In this study 1-D and 2-D gel electrophoresis-based protocols were used for the analysis of proteins in the TX-114 extract of<jats:italic>M. tuberculosis</jats:italic>membranes. Peptide mass mapping (using MALDI-TOF-MS, matrix assisted laser desorption/ionization time of flight mass spectrometry) of 116 samples led to the identification of 105 proteins, 9 of which were new to the<jats:italic>M. tuberculosis</jats:italic>proteome. Functional orthologues of 73 of these proteins were also present in<jats:italic>Mycobacterium leprae</jats:italic>, suggesting their relative importance. Bioinformatics predicted that as many as 73 % of the proteins had a hydrophobic disposition. 1-D gel electrophoresis revealed more hydrophobic/transmembrane and basic proteins than 2-D gel electrophoresis. Identified proteins fell into the following major categories: protein synthesis, cell wall biogenesis/architecture and conserved hypotheticals/unknowns. To identify immunodominant proteins of the detergent phase (DP), 14 low-molecular-mass fractions prepared by continuous-elution gel electrophoresis were subjected to T cell activation assays using blood samples from BCG-vaccinated healthy donors from a tuberculosis endemic area. Analysis of the responses (cell proliferation and IFN-<jats:italic>γ</jats:italic>production) showed that the immunodominance of certain DP fractions was most probably due to ribosomal proteins, which is consistent with both their specificity for mycobacteria and their abundance. Other membrane-associated proteins, including transmembrane proteins/lipoproteins and ESAT-6, did not appear to contribute significantly to the observed T cell responses.</jats:p> |
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| Umfang: | 2411-2419 |
| ISSN: |
1465-2080
1350-0872 |
| DOI: | 10.1099/mic.0.27799-0 |