Hiwasa T, Sawada T, Tanaka K, Chiba T, Tanaka T, Kominami E, Katunuma N, Sakiyama S
Division of Biochemistry, Chiba Cancer Center Research Institute, Japan.
Biomed Biochim Acta. 1991;50(4-6):579-85.
Subcellular localisation of Ha-ras gene products (p21) and cathepsin L was investigated by immunocytochemical studies. Western blotting analysis by using v-Ha-ras p21 deletion-insertion mutants indicated that NCC-RAS-001 and -004 monoclonal antibodies recognize epitopes within amino acids 126-140 and 146-165, respectively, of Ha-ras p21. Indirect immunofluorescent studies using these antibodies showed that ras p21 is distributed in patch-shaped structures in the cytoplasm. Cathepsin L whose activity is effectively inhibited by p21s in vitro was found in the regions where p21s are concentrated. These p21-concentrated regions may be derived from endosome-like vesicles because they are also labeled with Lucifer Yellow. The co-localization of p21 and cathepsin L in cytoplasmic vesicles was also confirmed by electron microscopic immunocytochemistry.
通过免疫细胞化学研究对Ha-ras基因产物（p21）和组织蛋白酶L进行亚细胞定位。使用v-Ha-ras p21缺失-插入突变体的蛋白质印迹分析表明，NCC-RAS-001和-004单克隆抗体分别识别Ha-ras p21氨基酸126-140和146-165内的表位。使用这些抗体的间接免疫荧光研究表明，ras p21分布于细胞质中的斑块状结构中。在p21集中的区域发现了其活性在体外被p21有效抑制的组织蛋白酶L。这些p21集中区域可能源自内体样小泡，因为它们也被路西法黄标记。电子显微镜免疫细胞化学也证实了p21和组织蛋白酶L在细胞质小泡中的共定位。
