Real-time monitoring of the PDE2 activity of live cells: hormone-stimulated
cAMP hydrolysis is faster than hormone-stimulated cAMP synthesis
V. Nikolaev, S. Gambaryan, S. Engelhardt, U. Walter, and M. Lohse. J Biol Chem, 280 (3):
1716-9(January 2005)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..
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.
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.
%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
}