Genetically encoded FRET probe for PKC activity based on pleckstrin
Genetically encoded FRET probe for PKC activity based on pleckstrin
复制标题
DOI：
10.1021/ja0460155
复制
发表时间：
2004-09-29
影响因子：
15
通讯作者：
Schultz, C
中科院分区：
文献类型：
--
作者：
Schleifenbaum, A;Stier, G;Schultz, C
我们开发了一种检测活细胞中蛋白激酶C （PKC）活性的探针。探针是基于一个片段的pleckstring封闭的两个FRET-capable荧光团。体外和体内PKC活性的调节可靠地跟随FRET的变化。探针对磷酸酯激活PKC反应迅速。当使用PKC抑制剂时，FRET变化是可逆的。使用组胺或缓激肽刺激细胞信号通路以生理相关的方式触发PKC。
We developed a probe for investigating protein kinase C (PKC) activity in living cells. The probe is based on a fragment of pleckstrin enclosed by two FRET-capable fluorophores. PKC activity modulation was reliably followed by FRET change in vitro and in vivo. The probe responds quickly to PKC activation by phorbol ester. FRET changes were reversible when PKC inhibitors were administered. Stimulation of cellular signaling pathways using histamine or bradykinin triggered PKC in a physiologically relevant way.