Microtubule-depolymerizing agents inhibit asialo-orosomucoid delivery to lysosomes but not its endocytosis or degradation in isolated rat hepatocytes.
Microtubule-depolymerizing drugs, such as colchicine, vinblastine sulfate, colcemide and podophyllotoxin, cause an apparent inhibition of the ability of rat hepatocytes to degrade asialo-orosomucoid. However, the binding of asialo-orosomucoid to the cell surface at 0 degrees C, the endocytosis of pre-bound glycoprotein at 37 degrees C, and the dissociation of internal receptor-glycoprotein complexes are unaffected by these microtubule drugs. Receptor recycling is slowed but still occurs, although degradation is blocked. The rate of degradation is decreased by low concentrations of drugs. (For example, 0.25 microM vinblastine sulfate, colchicine and colcemide inhibited 93%, 79% and 26%, respectively.) Neither beta- nor gamma-lumicolchicine affected any of the processes examined. The degree of inhibition with colchicine could be enhanced by a brief treatment of the cells at low temperature to depolymerize microtubules. However, if cells were allowed to endocytose asialo-orosomucoid at 37 degrees C prior to addition of the microtubule drug, then the inhibition of protein degradation was greatly reduced. The decrease in the inhibition of degradation was proportional to the amount of time that cells were exposed to asialoglycoprotein before addition of the drug. The results indicate that the segregation of protein from receptor after they dissociate and/or the subsequent translocation of internalized asialoglycoprotein from the cell perimeter to the lysosomal region requires intact microtubules.
D000818:Animals; D001212:Asialoglycoproteins; D003078:Colchicine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D004705:Endocytosis / Q000187:drug effects*; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D009961:Orosomucoid / Q000031:analogs & derivatives* / Q000378:metabolism; D011034:Podophyllotoxin / Q000494:pharmacology; D051381:Rats; D014747:Vinblastine / Q000494:pharmacology*
D001212:Asialoglycoproteins; D009961:Orosomucoid; C017030:asialoorosomucoid; D014747:Vinblastine; D011034:Podophyllotoxin; D003078:Colchicine