Detecting cAMP-induced Epac activation by fluorescence resonance energy transfer: Epac as a novel cAMP indicator Export

@article{citeulike:2015, abstract = {Epac1 is a guanine nucleotide exchange factor for Rap1 that is activated by direct binding of cAMP. In vitro studies suggest that cAMP relieves the interaction between the regulatory and catalytic domains of Epac. Here, we monitor Epac1 activation in vivo by using a CFP−Epac−YFP fusion construct. When expressed in mammalian cells, CFP−Epac−YFP shows significant fluorescence resonance energy transfer (FRET). FRET rapidly decreases in response to the cAMP-raising agents, whereas it fully recovers after addition of cAMP-lowering agonists. Thus, by undergoing a cAMP-induced conformational change, CFP−Epac−YFP serves as a highly sensitive cAMP indicator in vivo. When compared with a protein kinase A (PKA)-based sensor, Epac-based cAMP probes show an extended dynamic range and a better signal-to-noise ratio; furthermore, as a single polypeptide, CFP−Epac−YFP does not suffer from the technical problems encountered with multisubunit PKA-based sensors. These properties make Epac-based FRET probes the preferred indicators for monitoring cAMP levels in vivo.}, author = {Ponsioen, Bas and Zhao, Jun and Riedl, Jurgen and Zwartkruis, Fried and van der Krogt, Gerard and Zaccolo, Manuela and Moolenaar, Wouter H. and Bos, Johannes L. and Jalink, Kees}, citeulike-article-id = {2015}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/sj.embor.7400290}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/15550931}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=15550931}, day = {26}, doi = {10.1038/sj.embor.7400290}, journal = {EMBO Reports}, keywords = {camp, epac, fret}, month = {November}, number = {12}, posted-at = {2006-09-19 12:55:56}, priority = {2}, title = {Detecting cAMP-induced Epac activation by fluorescence resonance energy transfer: Epac as a novel cAMP indicator}, url = {http://dx.doi.org/10.1038/sj.embor.7400290}, volume = {5}, year = {2004} }

TY - JOUR ID - citeulike:2015 L3 - citeulike-article-id:2015 N2 - Epac1 is a guanine nucleotide exchange factor for Rap1 that is activated by direct binding of cAMP. In vitro studies suggest that cAMP relieves the interaction between the regulatory and catalytic domains of Epac. Here, we monitor Epac1 activation in vivo by using a CFP−Epac−YFP fusion construct. When expressed in mammalian cells, CFP−Epac−YFP shows significant fluorescence resonance energy transfer (FRET). FRET rapidly decreases in response to the cAMP-raising agents, whereas it fully recovers after addition of cAMP-lowering agonists. Thus, by undergoing a cAMP-induced conformational change, CFP−Epac−YFP serves as a highly sensitive cAMP indicator in vivo. When compared with a protein kinase A (PKA)-based sensor, Epac-based cAMP probes show an extended dynamic range and a better signal-to-noise ratio; furthermore, as a single polypeptide, CFP−Epac−YFP does not suffer from the technical problems encountered with multisubunit PKA-based sensors. These properties make Epac-based FRET probes the preferred indicators for monitoring cAMP levels in vivo. IS - 12 JF - EMBO Reports TI - Detecting cAMP-induced Epac activation by fluorescence resonance energy transfer: Epac as a novel cAMP indicator VL - 5 KW - camp KW - epac KW - fret AU - Ponsioen, Bas AU - Zhao, Jun AU - Riedl, Jurgen AU - Zwartkruis, Fried AU - van der Krogt, Gerard AU - Zaccolo, Manuela AU - Moolenaar, Wouter AU - Bos, Johannes AU - Jalink, Kees PY - 2004/11/26/ UR - http://dx.doi.org/10.1038/sj.embor.7400290 ER -