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Expression, immunochemical characterization and localization of the Mycobacterium tuberculosis protein p27
V Le Moigne1, G Robreau, C Borot
1Laboratoire Universitaire de Microbiologie Appliquée de Quimper (LUMAQ), UBO, 2 rue de l'Université, Quimper 29334 cedex, France.
Abstract:
Rv2108 is a gene of the PPE family of Mycobacterium tuberculosis specific for this bacterial complex and that may encode a putative protein p27. This gene was amplified, inserted into bacterial vectors, sequenced, and expressed as a recombinant protein. Specific antibodies to this protein were generated and used for immunochemical characterization and cellular localization. Mass spectrometric analysis of the expressed protein revealed a molecule that corresponded to the p27 putative protein. The expressed protein was immunologically active, and reacted with antibodies from tuberculosis patient sera. Specific immunoblot analysis confirmed the presence of the p27 antigen in Mycobacterium bovis BCG strain and in human clinical isolates of M. tuberculosis, but not in other mycobacteria tested. Western blot and immunoelectron microscopic analysis of BCG strain indicated that the p27 protein is localized in the membrane of the cell. The specific expression of the p27 protein in the M. tuberculosis complex could provide a novel specific complimentary diagnostic test for the presence of and infection with M. tuberculosis.
Insights
Researchers identified a specific protein, p27, from the Mycobacterium tuberculosis complex. This protein shows potential as a novel diagnostic marker for tuberculosis detection in patients.
Area of Science:
- Microbiology
- Immunology
- Molecular Biology
Background:
- The PPE gene family in Mycobacterium tuberculosis is crucial for its survival and pathogenesis.
- Rv2108, a member of the PPE family, encodes a putative protein p27, specific to the M. tuberculosis complex.
Purpose of the Study:
- To characterize the Rv2108 gene product (p27) and evaluate its potential as a diagnostic marker for tuberculosis.
Main Methods:
- Gene amplification, cloning, sequencing, and recombinant protein expression of Rv2108.
- Generation of specific antibodies for immunochemical characterization and cellular localization.
- Mass spectrometry, immunoblot analysis, Western blot, and immunoelectron microscopy were employed.
Main Results:
- The expressed protein was confirmed as p27 and demonstrated immunological activity.
- p27 antigen was detected in Mycobacterium bovis BCG and clinical M. tuberculosis isolates, but not in other mycobacteria.
- Immunoelectron microscopy localized p27 to the cell membrane of BCG.
Conclusions:
- The p27 protein is specifically expressed within the M. tuberculosis complex.
- This specific expression suggests p27's potential as a novel diagnostic target for tuberculosis infections.
