Inhibitory effect of ethanol and colchicine on the intracellular processing of beta-glucuronidase which occurs in the Golgi complex.
Inhibitory effect of ethanol and colchicine on the intracellular processing of beta-glucuronidase which occurs in the Golgi complex.
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乙醇和秋水仙碱对高尔基复合体中发生的β-葡萄糖醛酸酶的细胞内加工有抑制作用。
DOI：
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发表时间：
1995
影响因子：
2
通讯作者：
M. Himeno
中科院分区：
文献类型：
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作者：
Y. Nishimura;K. Kato;K. Oda;M. Himeno
为了研究乙醇或秋水仙碱对细胞内高尔基复合体中溶酶体β -葡萄糖醛酸酶的蛋白水解过程的影响，从乙醇或秋水仙碱处理的大鼠肝脏中分离出三个高尔基亚段，GF-1， GF-2和GF-3，并检测提取的高尔基β -葡萄糖醛酸酶在聚丙烯酰胺凝胶上的电泳模式。从给药大鼠中分离出的高尔基体亚段比从对照大鼠中分离出的高尔基体得率更高。这三个亚组分的酶学表征表明，它们没有受到其他亚细胞结构（如质膜、微粒体或溶酶体）的明显污染，也没有观察到药物的抑制作用。另一方面，在秋水仙碱处理的大鼠中检测到高尔基复合物标记酶半乳糖转移酶活性受到抑制。高尔基β -葡萄糖醛酸酶在聚丙烯酰胺凝胶上的电泳图谱显示有一条主带移动到与对照组溶酶体酶型相同的位置。相反，在乙醇和秋水草碱处理的大鼠中，高尔基β -葡萄糖醛酸酶被发现有两个主要的酶活性染色带，这是由微粒体和溶酶体型酶的混合物引起的。这些结果表明，在从微体型酶转化为溶酶体型酶的过程中，翻译后修饰步骤明显受到抑制。综上所述，这些发现表明，乙醇或秋水仙碱给药对以溶酶体为靶点的高尔基β -葡萄糖醛酸酶的细胞内翻译后修饰有抑制作用。
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.