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 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.
journal_name
Infect Immunjournal_title
Infection and immunityauthors
Harboe M,Wiker HG,Ulvund G,Lund-Pedersen B,Andersen AB,Hewinson RG,Nagai Sdoi
10.1128/IAI.66.1.289-296.1998subject
Has Abstractpub_date
1998-01-01 00:00:00pages
289-96issue
1eissn
0019-9567issn
1098-5522journal_volume
66pub_type
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journal_title:Infection and immunity
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journal_title:Infection and immunity
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更新日期:1990-06-01 00:00:00
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journal_title:Infection and immunity
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journal_title:Infection and immunity
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更新日期:1998-05-01 00:00:00
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