Burge V, Mainferme F, Wattiaux R
Laboratoire de Chimie Physiologique, Facultés Universitaries Notre Dame de la Paix, Namur, Belgium.
Biochem J. 1991 May 1;275 ( Pt 3)(Pt 3):797-800. doi: 10.1042/bj2750797.
Transport of the lysosomal enzyme cathepsin C was studied in Morris hepatoma 7777 cells. Subcellular fractions obtained after isopyenic centrifugation in sucrose gradients of labelled cell homogenates were sequentially extracted by hypo-osmotic shock, Na2CO3 and Triton X-100. Polypeptides related to cathepsin C were immunoprecipitated and analysed by SDS/PAGE and fluorography. At early times after synthesis and for up to 60 min, precursor polypeptides of cathepsin C are distributed in endoplasmic reticulum and Golgi fractions, in membrane-associated form, as Triton X-100 is necessary for their extraction. At 2 h and later after synthesis, intermediate and mature forms of the enzyme can be totally extracted by hypo-osmotic shock from gradient fractions corresponding to the lysosomes of Morris hepatoma 7777 cells.
在莫里斯肝癌7777细胞中研究了溶酶体酶组织蛋白酶C的转运。将标记的细胞匀浆在蔗糖梯度中进行等密度离心后获得的亚细胞组分，依次通过低渗休克、碳酸钠和曲拉通X-100进行提取。与组织蛋白酶C相关的多肽通过免疫沉淀法进行沉淀，并通过SDS/聚丙烯酰胺凝胶电泳和荧光自显影进行分析。在合成后的早期以及长达60分钟内，组织蛋白酶C的前体多肽以膜结合形式分布在内质网和高尔基体组分中，因为曲拉通X-100对于它们的提取是必需的。在合成后2小时及更晚的时间，该酶的中间形式和成熟形式可以通过低渗休克从对应于莫里斯肝癌7777细胞溶酶体的梯度组分中完全提取出来。
