An inhibitory effect of a protein kinase inhibitor, staurosporine, on delivery of endocytosed asialoglycoprotein to lysosome in monolayer culture of rat hepatocytes.
An inhibitor of protein kinases, staurosporine (ssp), was found to affect the endocytic pathway of asialoglycoproteins subsequent to endocytosis in monolayer cultures of rat hepatocytes. The effect of 5 or 10 microM staurosporine on the internalization of a synthetic ligand (galBSA-HRP: bovine serum albumin exposing galactose, horseradish peroxidase conjugates) prebound to the cell surface was minimal. The presence of 5, 7, or 10 microM ssp during a 1-h chase period resulted in the ligand remaining in a low density (1.04-1.05 g/ml), nonlysosomal subcellular fraction in a Percoll gradient. The ligand, arrested by 7 microM ssp, was further processed to the lysosome during subsequent incubation in the absence of ssp. Cells maintained the ability to internalize ligand at 37 degrees C for 1 h in the presence of these concentrations of ssp. During a 1-h continuous uptake of 0-50 micrograms/ml nonlabeled ligand, the presence of 7 microM ssp did not cause any decrease in the amount of asialoglycoprotein receptor at the cell surface, which indicates receptor recycling occurred normally. These results suggest a possible involvement of protein kinase(s), which can be inhibited by ssp, in the delivery of endocytosed ligand to the lysosome, but not in ligand endocytosis and receptor recycling.
Ohashi
M
M
Department of Biophysics, Faculty of Science, Kyoto University, Japan.
Ohnishi
S
S
D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D001692:Biological Transport / Q000187:drug effects; D002458:Cell Fractionation; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D004705:Endocytosis / Q000187:drug effects*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009961:Orosomucoid / Q000031:analogs & derivatives* / Q000378:metabolism; D047428:Protein Kinase Inhibitors; D051381:Rats; D011919:Rats, Inbred Strains; D011971:Receptors, Immunologic / Q000378:metabolism*; D019311:Staurosporine
D000470:Alkaloids; D037263:Asialoglycoprotein Receptor; D001212:Asialoglycoproteins; D009961:Orosomucoid; D047428:Protein Kinase Inhibitors; D011971:Receptors, Immunologic; C017030:asialoorosomucoid; D019311:Staurosporine