Inhibitory effect of ethanol and colchicine on the intracellular processing of beta-glucuronidase which occurs in the Golgi complex.
To investigate the effect of ethanol or colchicine on the intracellular proteolytic processing of lysosomal beta-glucuronidase, which is considered to occur in the Golgi complex in the intracellular sorting pathway, three rat liver Golgi subfractions, GF-1, GF-2, and GF-3, were isolated from ethanol- or colchicine-treated rats, and the electrophoretic patterns of the extracted Golgi beta-glucuronidase on polyacrylamide gel were examined. The isolated Golgi subfractions from the drug-treated rats gave a better yield of fraction than that from the control rats. The enzymatic characterization of these three subfractions showed no significant contamination by other subcellular structures such as plasma membranes, microsomes, or lysosomes, and no inhibitory effect of the drugs was observed. On the other hand, suppressed galactosyltransferase activity, a marker enzyme of the Golgi complex, was detected in the colchicine-treated rats. The electrophoretic pattern of Golgi beta-glucuronidase on polyacrylamide gel revealed one major band which moved to the same position as the lysosomal enzyme type in the control rats. In contrast, in the ethanol- and colchicine-treated rats, Golgi beta-glucuronidase was found to have two major bands stained for enzyme activity resulting from a mixture of microsomal- and lysosomal-type enzymes. These results suggested that the post-translational modification step, during conversion from a microsomal-type enzyme to a lysosomal-type enzyme, was apparently inhibited. Taken together, these findings indicated that ethanol or colchicine administration to rats caused an inhibitory effect on the intracellular post-translational modification of Golgi beta-glucuronidase destined for targeting to the lysosomes.
Nishimura
Y
Y
Division of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Kato
K
K
Oda
K
K
Himeno
M
M
D000818:Animals; D003078:Colchicine / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol / Q000494:pharmacology*; D005966:Glucuronidase / Q000378:metabolism*; D006056:Golgi Apparatus / Q000187:drug effects / Q000201:enzymology*; D007525:Isoelectric Focusing; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology
D000431:Ethanol; D005966:Glucuronidase; D003078:Colchicine