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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. Russelldagger

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.

dagger 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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