Zaccolo M, De Giorgi F, Cho C Y, Feng L, Knapp T, Negulescu P A, Taylor S S, Tsien R Y, Pozzan T
Department of Experimental Biomedical Sciences, University of Padova, 35121 Padova, Italy.
Nat Cell Biol. 2000 Jan;2(1):25-9. doi: 10.1038/71345.
Cyclic AMP controls several signalling cascades within cells, and changes in the amounts of this second messenger have an essential role in many cellular events. Here we describe a new methodology for monitoring the fluctuations of cAMP in living cells. By tagging the cAMP effector protein kinase A with two suitable green fluorescent protein mutants, we have generated a probe in which the fluorescence resonance energy transfer between the two fluorescent moieties is dependent on the levels of cAMP. This new methodology opens the way to the elucidation of the biochemistry of cAMP in vivo.
环磷酸腺苷（cAMP）控制细胞内的多个信号级联反应，这种第二信使数量的变化在许多细胞事件中起着至关重要的作用。在此，我们描述了一种监测活细胞中cAMP波动的新方法。通过用两个合适的绿色荧光蛋白突变体标记cAMP效应蛋白激酶A，我们构建了一种探针，其中两个荧光部分之间的荧光共振能量转移取决于cAMP的水平。这种新方法为阐明体内cAMP的生物化学过程开辟了道路。
