Quantitative measurement of cAMP concentration using an exchange protein directly activated by a cAMP-based FRET-sensor.

ABSTRACT: Förster resonance energy transfer (FRET)-based biosensors for the quantitative analysis of intracellular signaling, including sensors for monitoring cyclic adenosine monophosphate (cAMP), are of increasing interest. The measurement of the donor/acceptor emission ratio in tandem biosensors excited at the donor excitation wavelength is a commonly used technique. A general problem, however, is that this ratio varies not only with the changes in cAMP concentration but also with the changes of the ionic environment or other factors affecting the folding probability of the fluorophores. Here, we use a spectral FRET analysis on the basis of two excitation wavelengths to obtain a reliable measure of the absolute cAMP concentrations with high temporal and spatial resolution by using an "exchange pr

SUBMITTER: Salonikidis PS 

PROVIDER: S-EPMC2586560 | biostudies-literature | 2008 Dec

REPOSITORIES: biostudies-literature