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@ARTICLE{Gruteser:909072,
author = {Gruteser, Nadine and Baumann, Arnd},
title = {{E}xamination of {I}ntracellular {GPCR}-{M}ediated
{S}ignaling with {H}igh {T}emporal {R}esolution},
journal = {International journal of molecular sciences},
volume = {23},
number = {15},
issn = {1422-0067},
address = {Basel},
publisher = {Molecular Diversity Preservation International},
reportid = {FZJ-2022-02990},
pages = {8516 -},
year = {2022},
note = {Personal costs (N.G.) and consumables of the study were
partly funded by the German Federal Ministry of Education
and Research (BMBF) within the project Molecular Interaction
Engineering, FKZ 031A095A (awarded to A.B.). Publication
costs were funded by the Deutsche Forschungsgemeinschaft
(DFG, German Research Foundation)},
abstract = {The GTP-binding protein-coupled receptors (GPCRs) play
important roles in physiology and neuronal signaling. More
than a thousand genes, excluding the olfactory receptors,
have been identified that encode these integral membrane
proteins. Their pharmacological and functional properties
make them fascinating targets for drug development, since
various disease states can be treated and overcome by
pharmacologically addressing these receptors and/or their
downstream interacting partners. The activation of the GPCRs
typically causes transient changes in the intracellular
second messenger concentrations as well as in membrane
conductance. In contrast to ion channel-mediated electrical
signaling which results in spontaneous cellular responses,
the GPCR-mediated metabotropic signals operate at a
different time scale. Here we have studied the kinetics of
two common GPCR-induced signaling pathways: (a) Ca2+ release
from intracellular stores and (b) cyclic adenosine
monophosphate (cAMP) production. The latter was monitored
via the activation of cyclic nucleotide-gated (CNG) ion
channels causing Ca2+ influx into the cell. Genetically
modified and stably transfected cell lines were established
and used in stopped-flow experiments to uncover the
individual steps of the reaction cascades. Using two
homologous biogenic amine receptors, either coupling to Go/q
or Gs proteins, allowed us to determine the time between
receptor activation and signal output. With ~350 ms, the
release of Ca2+ from intracellular stores was much faster
than cAMP-mediated Ca2+ entry through CNG channels (~6 s).
The measurements with caged compounds suggest that this
difference is due to turnover numbers of the GPCR downstream
effectors rather than the different reaction cascades, per
se},
cin = {IBI-1},
ddc = {540},
cid = {I:(DE-Juel1)IBI-1-20200312},
pnm = {5241 - Molecular Information Processing in Cellular Systems
(POF4-524)},
pid = {G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)16},
pubmed = {35955656},
UT = {WOS:000839069100001},
doi = {10.3390/ijms23158516},
url = {https://juser.fz-juelich.de/record/909072},
}