PMID: 9423870Jan 10, 1998Paper

MPB70 and MPB83 as indicators of protein localization in mycobacterial cells

Infection and Immunity
M HarboeS Nagai

Abstract

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 conta...Continue Reading

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Citations

Nov 13, 2004·FEMS Microbiology Reviews·Iain C Sutcliffe, Dean J Harrington
Aug 22, 2006·Scandinavian Journal of Immunology·H G WikerA M D Riise
May 15, 2009·Scandinavian Journal of Immunology·H G Wiker
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Jul 5, 2011·Proteomics·Gustavo A de Souza, Harald G Wiker
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Feb 27, 2007·Microbiology·Mandana RezwanPeter Sander
Oct 19, 2012·Virulence·Marina A ForrelladFabiana Bigi
Feb 3, 2000·Tubercle and Lung Disease : the Official Journal of the International Union Against Tuberculosis and Lung Disease·M Daffé, G Etienne

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