Subcellular distribution of protein kinase C of GH3 cells: quantitation and characterization by polyacrylamide gel electrophoresis.
Subcellular distribution of protein kinase C of GH3 cells: quantitation and characterization by polyacrylamide gel electrophoresis.
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GH3 细胞蛋白激酶 C 的亚细胞分布：聚丙烯酰胺凝胶电泳定量和表征。
DOI：
10.1016/0003-9861(85)90816-1
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发表时间：
1985
影响因子：
3.9
通讯作者：
U. Eppenberger
中科院分区：
文献类型：
--
作者：
D. Fabbro;R. Jungmann;U. Eppenberger
非变性条件下，用聚丙烯酰胺凝胶电泳法(PAGE)定量测定细胞内钙和磷脂依赖性蛋白激酶(PKC)活性。用该方法可以准确定量小于50μg的胞浆蛋白的蛋白激酶C活性。PAGE后细胞内PKC活性的恢复量与DEAE-纤维素层析法所得的活性相当。在2 mMEGTA/EDTA存在下的GH3细胞匀浆显示，细胞总PKC活性的80%存在于胞浆中。然而，在细胞破碎后亚细胞分离和EDTA/EGTA提取亚细胞部分的过程中省略了离子螯合剂，结果表明，总PKC的80%存在于溶酶体-线粒体和微粒体提取物中。对PKC活性的详细分析表明，胞内PKC与EDTA/EGTA从亚细胞组分增溶的PKC是一致的。总之，GH3细胞似乎含有一种表观分子量为90,000的PKC，它似乎通过钙依赖机制与膜联系在一起。
Quantitative estimation of cytosolic Ca2+- and phospholipid-dependent protein kinase (PKC) activity was performed by polyacrylamide gel electrophoresis under nondenaturating conditions (PAGE). With this method less than 50 μg of cytosol protein can be accurately quantitated for PKC activity. The amount of cytosolic PKC activity recovered after PAGE was comparable to the amount obtained by DEAE-cellulose chromatography. Homogenization of GH3 cells in the presence of 2 mmEGTA/EDTA revealed that 80% of the total cellular PKC activity resided in the cytosol. However, omission of the ion chelator during cell disruption followed by subcellular fractionation and extraction of subcellular fractions by EDTA/EGTA showed that 80% of the total PKC was found in the lysosomal-mitochondrial and microsomal extracts. Detailed analysis of PKC activities demonstrated that cytosolic PKC was identical with the PKC solubilized by EDTA/EGTA from subcellular fractions. In conclusion, GH3 cells appear to contain one species of PKC with an apparent molecular weight of 90,000 which seems to be associated with membranes via a calcium-dependent mechanism (or mechanisms).
大鼠垂体肿瘤细胞中促甲状腺素释放激素受体复合物的快速温度依赖性转化。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hinkle,PM;Kinsella,PA
通讯作者：
Kinsella,PA
促进肿瘤的佛波酯影响大鼠垂体细胞催乳素和生长激素的产生。
影响因子：
4.8
作者：
Osborne,R;TashjianJr,AH
通讯作者：
TashjianJr,AH