Kuettel M R, Schwoch G, Jungmann R A
Cell Biol Int Rep. 1984 Nov;8(11):949-57. doi: 10.1016/0309-1651(84)90193-0.
We have applied the indirect colloidal immunogold technique to examine the ultrastructural localization of the catalytic subunit C and the regulatory subunits RI and RII of cyclic AMP-dependent protein kinase in rat hepatocyte nuclei before and after glucagon or dibutyryl cyclic AMP administration. The technique allowed the identification and localization of all three subunits in hepatocyte nuclei. Morphometric quantitation of the relative staining density of nuclear subunits indicated an increase of immunogold staining of nuclear catalytic subunit but not of the regulatory subunits after glucagon or dibutyryl cyclic AMP stimulation. The increase of catalytic subunit occurred in a biphasic manner with peak levels 2-30 min and 90-150 min after stimulation. Our experiments represent the first reported use of the immunogold procedure to identify and localize protein kinase subunits in the nucleus.
我们应用间接胶体免疫金技术，检测了在给予胰高血糖素或二丁酰环磷酸腺苷前后，大鼠肝细胞核中环磷酸腺苷依赖性蛋白激酶催化亚基C以及调节亚基RI和RII的超微结构定位。该技术能够鉴定并定位肝细胞核中的所有三个亚基。对核亚基相对染色密度的形态计量学定量分析表明，在胰高血糖素或二丁酰环磷酸腺苷刺激后，核催化亚基的免疫金染色增加，而调节亚基的免疫金染色未增加。催化亚基的增加呈双相性，在刺激后2 - 30分钟和90 - 150分钟达到峰值水平。我们的实验是首次报道使用免疫金方法在细胞核中鉴定和定位蛋白激酶亚基。
