Program in Cell Biology, University of California, San Francisco, San Francisco, CA, USA.
Department of Psychiatry and Behavioral Sciences, University of California, San Francisco, San Francisco, CA, USA.
Methods Mol Biol. 2022;2483:339-349. doi: 10.1007/978-1-0716-2245-2_21.
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.
通常通过细胞核中净蛋白含量或激酶活性的变化来推断 cAMP 依赖性蛋白激酶催化亚基的核内进入。以前直接评估催化亚基核内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，可以实时检测在生理范围内 cAMP 刺激下，以内源性水平表达的催化亚基在活细胞中的核内进入。
