This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Harboe, M.
Right arrow Articles by Nagai, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Harboe, M.
Right arrow Articles by Nagai, S.

 Previous Article  |  Next Article 

Infect. Immun., Jan 1998, 289-296, Vol 66, No. 1
Copyright © 1998, American Society for Microbiology

MPB70 and MPB83 as indicators of protein localization in mycobacterial cells

M Harboe, HG Wiker, G Ulvund, B Lund-Pedersen, AB Andersen, RG Hewinson and S Nagai
Institute of Immunology and Rheumatology, University of Oslo, Norway. morten.harboe@labmed.uio.no

Culture fluids after growth of Mycobacterium bovis BCG on Sauton medium contain actively secreted proteins and proteins released by bacterial lysis. BCG culture fluids and sonicates of Mycobacterium tuberculosis and Mycobacterium paratuberculosis were tested after separation by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The localization of marker proteins was determined by enzyme-linked immunosorbent assay and Western blotting with selected monoclonal antibodies of known specificities. Soluble secreted proteins (MPB64 and proteins of the antigen 85 complex) and three heat shock proteins (DnaK, GroEL, and GroES) were recovered in a single peak after gel filtration, indicating their occurrence as a free monomer in the culture fluid and cytosol, respectively. Other constituents eluted in two distinct peaks during gel filtration. The first peak corresponded to the void volume, indicating complex formation between several proteins or attachment to lipids in the surface layer or the cytoplasmic membrane; the second peak corresponded to the expected monomer size indicated by SDS-PAGE under conditions that separate proteins from each other during sample preparation. The two-peak group contained constituents with known lipid contents, the 19- and 38-kDa lipoproteins and lipoarabinomannan. The 26-kDa form of MPB83 behaved similarly. After extraction with Triton X-114, these constituents entered into the detergent phase, confirming the lipoprotein nature of 26-kDa MPB83. The MPB83 molecule was shown to be available on the surface of BCG Tokyo bacilli for reaction with monoclonal antibody MBS43 by flow cytometry.


This article has been cited by other articles:

  • Rezwan, M., Grau, T., Tschumi, A., Sander, P. (2007). Lipoprotein synthesis in mycobacteria. Microbiology 153: 652-658 [Abstract] [Full Text]  
  • Carr, M. D., Bloemink, M. J., Dentten, E., Whelan, A. O., Gordon, S. V., Kelly, G., Frenkiel, T. A., Hewinson, R. G., Williamson, R. A. (2003). Solution Structure of the Mycobacterium tuberculosis Complex Protein MPB70: FROM TUBERCULOSIS PATHOGENESIS TO INHERITED HUMAN CORNEAL DISEASE. J. Biol. Chem. 278: 43736-43743 [Abstract] [Full Text]  
  • Wiker, H. G., Lyashchenko, K. P., Aksoy, A. M., Lightbody, K. A., Pollock, J. M., Komissarenko, S. V., Bobrovnik, S. O., Kolesnikova, I. N., Mykhalsky, L. O., Gennaro, M. L., Harboe, M. (1998). Immunochemical Characterization of the MPB70/80 and MPB83 Proteins of Mycobacterium bovis. Infect. Immun. 66: 1445-1452 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»