Murakami Y, Uehara Y, Yamamoto C, Fukazawa H, Mizuno S
Department of Antibiotics, National Institute of Health, Tokyo, Japan.
Exp Cell Res. 1991 Aug;195(2):338-44. doi: 10.1016/0014-4827(91)90382-5.
Herbimycin A, which has been known to inactivate and degrade p60v-src tyrosine kinase, induced an elevated synthesis of a protein with a molecular size of 70 kDa in A431 human epidermoid carcinoma cells. This protein showed the same migration distance on SDS-polyacrylamide gel electrophoresis as that of the protein induced in the cells by heat shock treatment, and this 70-kDa protein was identified as a member of the heat shock protein 70 family (hsp70) through immunoprecipitation with anti-hsp72/73 antibody and partial digestion with V8 protease. The induced level of the 70-kDa protein was dependent on the length of period and the concentration of herbimycin A treatment. Cellular fractionation and indirect immunofluorescence analyses revealed that the 70-kDa protein induced by herbimycin A was localized in the cytoplasm, in contrast to the nuclear distribution of hsp70 induced by heat treatment. Induction of hsp70 by herbimycin A was also observed in several other cells, including HeLa S3 cells, chicken embryo fibroblasts, NIH3T3 cells, and Rous sarcoma virus-transformed NIH3T3 cells.
已知能使p60v-src酪氨酸激酶失活并降解的除草菌素A，可诱导A431人表皮样癌细胞中一种分子量为70 kDa的蛋白质合成增加。该蛋白质在SDS-聚丙烯酰胺凝胶电泳上的迁移距离与热休克处理诱导细胞产生的蛋白质相同，通过用抗hsp72/73抗体进行免疫沉淀和用V8蛋白酶部分消化，鉴定该70 kDa蛋白质为热休克蛋白70家族（hsp70）的成员。70 kDa蛋白质的诱导水平取决于除草菌素A处理的时间长度和浓度。细胞分级分离和间接免疫荧光分析表明，除草菌素A诱导的70 kDa蛋白质定位于细胞质中，这与热处理诱导的hsp70定位于细胞核不同。在包括HeLa S3细胞、鸡胚成纤维细胞、NIH3T3细胞和劳氏肉瘤病毒转化的NIH3T3细胞在内的其他几种细胞中也观察到除草菌素A诱导hsp70。
