We investigated the protein composition of J774-E clone macrophage phagosomes isolated at different stages of phagolysosome biogenesis. Phagosomes formed by internalizing antibody-coated Staphylococcus aureus for 3 min followed by chase for 0, 4, 9, or 15 min were isolated by density gradient centrifugation. Enrichment and purity of the phagosome preparations were quantitated by radiolabeled ligand recovery, enzyme markers, and electron microscopy. One-dimensional SDS-PAGE analyses of the isolated phagosomes revealed virtually identical protein compositions. However, Western blot analyses with antibodies directed against selected proteins of known itineraries along the endocytic pathway demonstrated distinct differences in phagosome protein compositions. Accumulating within the maturing phagosome were the 31-kD subunit of the vacuolar proton pump, cathepsin D,beta-glucuronidase, the cation dependent mannose 6-phosphate receptor, and LAMP-1. Decreasing within the maturing phagosome were the FcII receptor, the mannose receptor, and alpha-adaptin. These results indicate that although the macrophage phagosome's total protein composition changes little during phagolysosome formation, the maturing phagosome both receives and eliminates, possibly by protein recycling, specific membrane and sequestered proteins.
A Pitt, L S Mayorga, P D Stahl, A L Schwartz
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E Carrasco-Marín, C Alvarez-Domínguez, F Leyva-Cobián |
European Journal of Immunology | 1994 |
Biogenesis of phagolysosomes proceeds through a sequential series of interactions with the endocytic apparatus
M Desjardins, LA Huber, RG Parton, G Griffiths |
The Journal of Cell Biology | 1994 |
Calcium-dependent fusion among endosomes
LS Mayorga, W Berón, MN Sarrouf, MI Colombo, C Creutz, PD Stahl |
The Journal of biological chemistry | 1994 |
Molecular characterization of phagosomes
M Desjardins, JE Celis, G van Meer, H Dieplinger, A Jahraus, G Griffiths, LA Huber |
The Journal of biological chemistry | 1994 |
Association of Rap1a and Rap1b proteins with late endocytic/phagocytic compartments and Rap2a with the Golgi complex
V Pizon, M Desjardins, C Bucci, RG Parton, M Zerial |
Journal of cell science | 1994 |
Fusion between large phagocytic vesicles: targeting of yeast and other particulates to phagolysosomes that shelter the bacterium Coxiella burnetii or the protozoan Leishmania amazonensis in Chinese hamster ovary cells
PS Veras, C de Chastellier, MF Moreau, V Villiers, M Thibon, D Mattei, M Rabinovitch |
Journal of cell science | 1994 |