Morphological localization of a major lysosomal membrane glycoprotein in the endocytic membrane system.
We have raised specific polyclonal immunoglobulin G (IgG) against a major lysosomal membrane sialoglycoprotein (LGP107) taken from rat liver and have prepared a conjugate of its Fab' fragment with horseradish peroxidase (HRP-anti LGP107 Fab') as a probe for the subcellular antigen. Electron immunocytochemistry in primary cultured rat hepatocytes showed that LGP107 resided primarily within lysosomes and was associated with luminal amorphous materials as well as limiting membranes. In addition, LGP107 was shown to be substantially distributed throughout the endocytic vacuolar system. The glycoprotein was found clustered in coated pits at the cell surface and localized along the surrounding membranes in endocytic vesicles. When cultured cells were exposed to HRP-anti LGP107 Fab', the antibody which was bound to its antigen within the coated pits was internalized via a system of endocytic vesicles and transported to lysosomes. During 20 min of incubation at 37 degrees C, the HRP tracer appeared at an early stage in small vesicles and moved progressively to larger vesicles, including multivesicular bodies. After 1 h, the tracer could be clearly seen in lysosomes heterogeneous in shape and size. The existence of LGP107 in endocytic compartments and the uptake of anti LGP107 antibody by hepatocytes were not blocked by prior treatment of the cells with cycloheximide and excess amounts of anti LGP107 IgG. These data suggest that LGP107 circulates between the cell surface and lysosomes through the endocytic membrane traffic in hepatocytes.
Furuno
K
K
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka.
Ishikawa
T
T
Akasaki
K
K
Yano
S
S
Tanaka
Y
Y
Yamaguchi
Y
Y
Tsuji
H
H
Himeno
M
M
Kato
K
K
D000818:Animals; D002478:Cells, Cultured; D006735:Horseradish Peroxidase; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology; D008247:Lysosomes / Q000032:analysis*; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D012795:Sialoglycoproteins / Q000032:analysis* / Q000276:immunology
D007140:Immunoglobulin Fab Fragments; D008562:Membrane Glycoproteins; D012795:Sialoglycoproteins; D006735:Horseradish Peroxidase