%0 Journal Article %1 Nikolaev2005 %A Nikolaev, V. O. %A Gambaryan, S. %A Engelhardt, S. %A Walter, U. %A Lohse, M. J. %D 2005 %J J Biol Chem %K AMP/*metabolism Activation Animals Atrial Cyclic Diester Energy Enzyme Factor/pharmacology Fluorescence Forskolin/pharmacology Glomerulosa/cytology/drug Hydrolases/*metabolism Hydrolysis Natriuretic Phosphoric Resonance Transfer Zona effects/enzymology %N 3 %P 1716-9 %T Real-time monitoring of the PDE2 activity of live cells: hormone-stimulated cAMP hydrolysis is faster than hormone-stimulated cAMP synthesis %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15557342 %V 280 %X Cyclic nucleotide phosphodiesterases (PDEs) are the enzymes that catalyze the hydrolysis of cAMP and cGMP, thereby restricting the activity of these second messengers in cells. A unique ability to shape gradients of cyclic nucleotides and compartmentalize their signaling implies a high potency and a rapid action of PDEs. However, it has not been demonstrated how fast PDEs can hydrolyze cAMP in a living system. Here we perform a real-time monitoring of PDE2 activity in aldosterone-producing adrenal cells using a recently developed genetically encoded, fluorescent cAMP sensor, which reveals enormously rapid kinetics of cAMP degradation. Activation of PDE2 results in a rapid decrease of intracellular cAMP from high micromolar to the sub-micromolar range within a few seconds. Moreover, the kinetics of atrial natriuretic peptide-stimulated PDE2 activity (measured as decline of cAMP) are much faster than the speed of ACTH and isoprenaline-induced cAMP-synthesis (measured as cAMP accumulation) in the cells, revealing high catalytic activity and fast action of PDEs in regulating cAMP signaling in a physiological system.

@article{Nikolaev2005, abstract = {Cyclic nucleotide phosphodiesterases (PDEs) are the enzymes that catalyze the hydrolysis of cAMP and cGMP, thereby restricting the activity of these second messengers in cells. A unique ability to shape gradients of cyclic nucleotides and compartmentalize their signaling implies a high potency and a rapid action of PDEs. However, it has not been demonstrated how fast PDEs can hydrolyze cAMP in a living system. Here we perform a real-time monitoring of PDE2 activity in aldosterone-producing adrenal cells using a recently developed genetically encoded, fluorescent cAMP sensor, which reveals enormously rapid kinetics of cAMP degradation. Activation of PDE2 results in a rapid decrease of intracellular cAMP from high micromolar to the sub-micromolar range within a few seconds. Moreover, the kinetics of atrial natriuretic peptide-stimulated PDE2 activity (measured as decline of cAMP) are much faster than the speed of ACTH and isoprenaline-induced cAMP-synthesis (measured as cAMP accumulation) in the cells, revealing high catalytic activity and fast action of PDEs in regulating cAMP signaling in a physiological system.}, added-at = {2010-12-14T18:12:02.000+0100}, author = {Nikolaev, V. O. and Gambaryan, S. and Engelhardt, S. and Walter, U. and Lohse, M. J.}, biburl = {https://www.bibsonomy.org/bibtex/22e45eb8a320ecf093e12ca9f12228838/pharmawuerz}, endnotereftype = {Journal Article}, interhash = {6a4e6135fddaf4815082614cff6a3e79}, intrahash = {2e45eb8a320ecf093e12ca9f12228838}, issn = {0021-9258 (Print) 0021-9258 (Linking)}, journal = {J Biol Chem}, keywords = {AMP/*metabolism Activation Animals Atrial Cyclic Diester Energy Enzyme Factor/pharmacology Fluorescence Forskolin/pharmacology Glomerulosa/cytology/drug Hydrolases/*metabolism Hydrolysis Natriuretic Phosphoric Resonance Transfer Zona effects/enzymology}, month = {Jan 21}, note = {Nikolaev, Viacheslav O Gambaryan, Stepan Engelhardt, Stefan Walter, Ulrich Lohse, Martin J Research Support, Non-U.S. Gov't United States The Journal of biological chemistry J Biol Chem. 2005 Jan 21;280(3):1716-9. Epub 2004 Nov 22.}, number = 3, pages = {1716-9}, shorttitle = {Real-time monitoring of the PDE2 activity of live cells: hormone-stimulated cAMP hydrolysis is faster than hormone-stimulated cAMP synthesis}, timestamp = {2010-12-14T18:22:03.000+0100}, title = {Real-time monitoring of the PDE2 activity of live cells: hormone-stimulated cAMP hydrolysis is faster than hormone-stimulated cAMP synthesis}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15557342}, volume = 280, year = 2005 }