Lysosomal membrane proteins of Amoeba proteus, as studied with monoclonal antibodies.
Monoclonal antibodies were prepared against lysosomal membrane proteins of amoebae and used to follow lysosome-phagosome fusion after induced phagocytosis. The specificity of antibodies was checked by indirect immunofluorescence microscopy, immunoelectron microscopy, and localization of the antigen in subcellular fractions. The antibody-recognized proteins started to appear on the membranes of phagolysosomes about 5 min after phagocytosis as detected by indirect immunofluorescence, and the intensity of fluorescence increased for up to 1 h. Results of injection experiments in which purified antibodies had been injected into living cells and probed by indirect fluorescence indicated that the antigens were located on the cytoplasmic side of the lysosomal membranes. Lysosomes fuse with phagosomes on the one hand but not with non-fusible vesicles such as symbiosomes on the other. The results support the view that a membrane component(s) of non-fusible vesicles somehow prevents lysosomes from fusing with them.
Choi
E Y
EY
Department of Zoology, University of Tennessee, Knoxville 37996.
Kim
K J
KJ
Jeon
K W
KW
D000135:Acid Phosphatase / Q000378:metabolism; D000656:Amoeba / Q000201:enzymology / Q000276:immunology / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000913:Antibodies, Protozoan / Q000276:immunology* / Q000302:isolation & purification; D005260:Female; D007425:Intracellular Membranes / Q000276:immunology / Q000378:metabolism*; D008247:Lysosomes / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D016253:Microscopy, Immunoelectron; D010587:Phagocytosis; D010588:Phagosomes / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D008565:Membrane Proteins; D000135:Acid Phosphatase