A genetically encoded, fluorescent indicator for cyclic AMP in living cells.
Zaccolo M., De Giorgi F., Cho CY., Feng L., Knapp T., Negulescu PA., Taylor SS., Tsien RY., Pozzan T.
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.