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Infection and Immunity, December 2000, p. 6997-7002, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Identification of Mycobacterial Surface Proteins
Released into Subcellular Compartments of Infected
Macrophages
Wandy L.
Beatty* and
David G.
Russell
Department of Molecular Microbiology,
Washington University School of Medicine, St. Louis, Missouri 63110
Received 9 June 2000/Returned for modification 14 August
2000/Accepted 5 September 2000
Considerable effort has focused on the identification of proteins
secreted from Mycobacterium spp. that contribute to the development of protective immunity. Little is known, however, about the
release of mycobacterial proteins from the bacterial phagosome and the
potential role of these molecules in chronically infected macrophages.
In the present study, the release of mycobacterial surface proteins
from the bacterial phagosome into subcellular compartments of infected
macrophages was analyzed. Mycobacterium bovis BCG was
surface labeled with fluorescein-tagged succinimidyl ester, an
amine-reactive probe. The fluorescein tag was then used as a marker for
the release of bacterial proteins in infected macrophages.
Fractionation studies revealed bacterial proteins within subcellular
compartments distinct from mycobacteria and mycobacterial phagosomes.
To identify these proteins, subcellular fractions free of bacteria were
probed with mycobacterium-specific antibodies. The fibronectin
attachment protein and proteins of the antigen 85-kDa complex were
identified among the mycobacterial proteins released from the bacterial phagosome.
*
Corresponding author. Mailing address: Department of
Molecular Microbiology, 660 South Euclid Ave., Washington University School of Medicine, St. Louis, MO 63110. Phone: (314) 362-4987. Fax:
(314) 362-1232. E-mail: beatty{at}borcim.wustl.edu.

Present address: Department of Microbiology and
Immunology, College of Veterinary Medicine, Cornell University, Ithaca,
NY
14853.
Infection and Immunity, December 2000, p. 6997-7002, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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