Abstract:
Nuclear entry of cAMP-dependent protein kinase catalytic subunits is typically inferred from changes in net protein amount or kinase activity in the nucleus. Previous methods to directly assess nuclear entry require kinase subunit overexpression and/or supraphysiological cAMP elevation. We describe a method to detect nuclear entry of catalytic subunits expressed at an endogenous level in living cells, stimulated by cAMP in a physiological range, and in real time.
摘要:
cAMP依赖性蛋白激酶催化亚基的核进入通常是根据核中净蛋白质量或激酶活性的变化来推断的。以前直接评估核进入的方法需要激酶亚基过表达和/或超生理cAMP升高。我们描述了一种方法来检测在活细胞中内源性水平表达的催化亚基的核进入,由生理范围内的cAMP刺激,和实时。