%0 Journal Article %1 Nikolaev2004 %A Nikolaev, V. O. %A Bunemann, M. %A Hein, L. %A Hannawacker, A. %A Lohse, M. J. %D 2004 %J J Biol Chem %K *Signal AMP/*metabolism Animals Cell Cricetinae Cyclic Fluorescence Humans Line Messenger Second Systems Transduction %N 36 %P 37215-8 %T Novel single chain cAMP sensors for receptor-induced signal propagation %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15231839 %V 279 %X cAMP is a universal second messenger of many G-protein-coupled receptors and regulates a wide variety of cellular events. cAMP exerts its effects via cAMP-dependent protein kinase (PKA), cAMP-gated ion channels, and two isoforms of exchange protein directly activated by cAMP (Epac). Here we report the development of novel fluorescent indicators for cAMP based on the cAMP-binding domains of Epac and PKA. Fluorescence resonance energy transfer between variants of green fluorescent protein (enhanced cyan fluorescent protein and enhanced yellow fluorescent protein) fused directly to the cAMP-binding domains was used to analyze spatial and temporal aspects of cAMP-signaling in different cells. In contrast to previously developed PKA-based indicators, these probes are comprised of only a single binding site lacking cooperativity, catalytic properties, and interactions with other proteins and thereby allow us to easily image free intracellular cAMP and rapid signaling events. Rapid beta-adrenergic receptor-induced cAMP signals were observed to travel with high speed ( approximately 40 microm/s) throughout the entire cell body of hippocampal neurons and peritoneal macrophages. The developed indicators could be ubiquitously applied to studying cAMP, its physiological role and spatio-temporal regulation.

@article{Nikolaev2004, abstract = {cAMP is a universal second messenger of many G-protein-coupled receptors and regulates a wide variety of cellular events. cAMP exerts its effects via cAMP-dependent protein kinase (PKA), cAMP-gated ion channels, and two isoforms of exchange protein directly activated by cAMP (Epac). Here we report the development of novel fluorescent indicators for cAMP based on the cAMP-binding domains of Epac and PKA. Fluorescence resonance energy transfer between variants of green fluorescent protein (enhanced cyan fluorescent protein and enhanced yellow fluorescent protein) fused directly to the cAMP-binding domains was used to analyze spatial and temporal aspects of cAMP-signaling in different cells. In contrast to previously developed PKA-based indicators, these probes are comprised of only a single binding site lacking cooperativity, catalytic properties, and interactions with other proteins and thereby allow us to easily image free intracellular cAMP and rapid signaling events. Rapid beta-adrenergic receptor-induced cAMP signals were observed to travel with high speed ( approximately 40 microm/s) throughout the entire cell body of hippocampal neurons and peritoneal macrophages. The developed indicators could be ubiquitously applied to studying cAMP, its physiological role and spatio-temporal regulation.}, added-at = {2010-12-14T18:12:02.000+0100}, author = {Nikolaev, V. O. and Bunemann, M. and Hein, L. and Hannawacker, A. and Lohse, M. J.}, biburl = {https://www.bibsonomy.org/bibtex/211792f424d47339c7325fe157e11afc4/pharmawuerz}, endnotereftype = {Journal Article}, interhash = {80f14b6174446283077d0e67aa561258}, intrahash = {11792f424d47339c7325fe157e11afc4}, issn = {0021-9258 (Print) 0021-9258 (Linking)}, journal = {J Biol Chem}, keywords = {*Signal AMP/*metabolism Animals Cell Cricetinae Cyclic Fluorescence Humans Line Messenger Second Systems Transduction}, month = {Sep 3}, note = {Nikolaev, Viacheslav O Bunemann, Moritz Hein, Lutz Hannawacker, Annette Lohse, Martin J Research Support, Non-U.S. Gov't United States The Journal of biological chemistry J Biol Chem. 2004 Sep 3;279(36):37215-8. Epub 2004 Jul 1.}, number = 36, pages = {37215-8}, shorttitle = {Novel single chain cAMP sensors for receptor-induced signal propagation}, timestamp = {2010-12-14T18:12:27.000+0100}, title = {Novel single chain cAMP sensors for receptor-induced signal propagation}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15231839}, volume = 279, year = 2004 }