Nishimura R N, Dwyer B E, de Vellis J, Clegg K B
Regeneration Research Laboratory, Veterans Affairs Hospital, Sepulveda, CA 91343.
Brain Res Mol Brain Res. 1992 Jan;12(1-3):203-8. doi: 10.1016/0169-328x(92)90085-p.
The heat shock response in a transformed astrocyte line was compared with nontransformed astrocytes. The synthesis of HSP 68, the major inducible heat shock protein (HSP 68) was induced by a non-lethal 45 degrees C, 10 min heat shock. Although the incorporation of [35S]methionine into HSP 68 suggested that similar amounts of protein were being synthesized after heat shock, Western immunoblotting demonstrated striking differences in the HSP immunostaining between the two cell types. By one- and 'two-dimensional gel electrophoresis the major 68 kDa heat shock protein (HSP 68) was similar in both cell types. However, HSP 68 from heat shocked, transformed astrocytes did not immunostain with the monoclonal antibody, C-92, which is specific for the major inducible heat shock protein of HeLa cells. In contrast HSP 68 from heat shocked, nontransformed astrocytes immunostained quite well. A polyclonal antibody raised against the inducible 72 kDa heat shock protein of HeLa cells immunostained the HSP 68 from both astrocytes and transformed astrocytes. Analysis of the mRNA from the two cell types after heat shock revealed two bands of approximately 2.5 and 2.8 kb in astrocytes but only a single 2.5 kb band in the heat shocked transformed astroglia. These results suggest that structural differences in the HSP 68 may be present in the transformed astrocytes compared to the normal astrocytes.
将转化的星形胶质细胞系中的热休克反应与未转化的星形胶质细胞进行比较。主要的可诱导热休克蛋白HSP 68的合成是由45摄氏度、10分钟的非致死性热休克诱导的。尽管[35S]甲硫氨酸掺入HSP 68表明热休克后合成的蛋白量相似，但Western免疫印迹显示两种细胞类型之间HSP免疫染色存在显著差异。通过一维和二维凝胶电泳，两种细胞类型中的主要68 kDa热休克蛋白（HSP 68）相似。然而，热休克后的转化星形胶质细胞中的HSP 68不能与单克隆抗体C-92发生免疫染色，该抗体对HeLa细胞的主要可诱导热休克蛋白具有特异性。相比之下，热休克后的未转化星形胶质细胞中的HSP 68免疫染色效果很好。针对HeLa细胞的可诱导72 kDa热休克蛋白产生的多克隆抗体对星形胶质细胞和转化星形胶质细胞中的HSP 68均进行了免疫染色。对热休克后两种细胞类型的mRNA分析显示，星形胶质细胞中有两条约2.5和2.8 kb的条带，而热休克后的转化星形胶质细胞中只有一条2.5 kb的条带。这些结果表明，与正常星形胶质细胞相比，转化星形胶质细胞中可能存在HSP 68的结构差异。
