Measurement of the millisecond activation switch of G protein-coupled
receptors in living cells
J. Vilardaga, M. Bunemann, C. Krasel, M. Castro, и M. Lohse. Nat Biotechnol, 21 (7):
807-12(июля 2003)Vilardaga, Jean-Pierre Bunemann, Moritz Krasel, Cornelius Castro,
Marian Lohse, Martin J Comparative Study Research Support, Non-U.S.
Gov't United States Nature biotechnology Nat Biotechnol. 2003 Jul;21(7):807-12.
Epub 2003 Jun 15..
Аннотация
Hormones and neurotransmitters transduce signals through G protein-coupled
receptors (GPCR). Despite their common signaling pathways, however,
the responses they elicit have different temporal patterns. To reveal
the molecular basis for these differences we have developed a generally
applicable fluorescence-based technique for real-time monitoring
of the activation switch of GPCRs in living cells. We used such direct
measurements to investigate the activation of the alpha(2A)-adrenergic
receptor (alpha(2A)AR; neurotransmitter) and the parathyroid hormone
receptor (PTHR; hormone) and observed much faster kinetics than expected:
approximately 40 ms for the alpha(2A)AR and approximately 1 s for
the PTHR. The different switch times are in agreement with the different
receptors' biological functions. Agonists and antagonists could rapidly
switch the receptors on or off, whereas a partial agonist caused
only a partial signal. This approach allows the comparison of agonist
and partial agonist intrinsic activities at the receptor level and
provides evidence for millisecond activation times of GPCRs.
Vilardaga, Jean-Pierre Bunemann, Moritz Krasel, Cornelius Castro,
Marian Lohse, Martin J Comparative Study Research Support, Non-U.S.
Gov't United States Nature biotechnology Nat Biotechnol. 2003 Jul;21(7):807-12.
Epub 2003 Jun 15.
%0 Journal Article
%1 Vilardaga2003
%A Vilardaga, J. P.
%A Bunemann, M.
%A Krasel, C.
%A Castro, M.
%A Lohse, M. J.
%D 2003
%J Nat Biotechnol
%K & 1/agonists/antagonists Binding Cell Clonidine/pharmacology Culture Fluorescence/*methods G-Protein-Coupled/agonists/antagonists GTP-Binding Hormone, Hormone/pharmacology Humans Kidney/chemistry/drug Line Parathyroid Protein Proteins/*chemistry/*metabolism Signal Spectrometry, Techniques/*methods Transduction/*physiology Type alpha-2/agonists/antagonists effects/metabolism inhibitors/*chemistry/*metabolism inhibitors/chemistry inhibitors/metabolism Receptor Adrenergic
%N 7
%P 807-12
%T Measurement of the millisecond activation switch of G protein-coupled
receptors in living cells
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12808462
%V 21
%X Hormones and neurotransmitters transduce signals through G protein-coupled
receptors (GPCR). Despite their common signaling pathways, however,
the responses they elicit have different temporal patterns. To reveal
the molecular basis for these differences we have developed a generally
applicable fluorescence-based technique for real-time monitoring
of the activation switch of GPCRs in living cells. We used such direct
measurements to investigate the activation of the alpha(2A)-adrenergic
receptor (alpha(2A)AR; neurotransmitter) and the parathyroid hormone
receptor (PTHR; hormone) and observed much faster kinetics than expected:
approximately 40 ms for the alpha(2A)AR and approximately 1 s for
the PTHR. The different switch times are in agreement with the different
receptors' biological functions. Agonists and antagonists could rapidly
switch the receptors on or off, whereas a partial agonist caused
only a partial signal. This approach allows the comparison of agonist
and partial agonist intrinsic activities at the receptor level and
provides evidence for millisecond activation times of GPCRs.
@article{Vilardaga2003,
abstract = {Hormones and neurotransmitters transduce signals through G protein-coupled
receptors (GPCR). Despite their common signaling pathways, however,
the responses they elicit have different temporal patterns. To reveal
the molecular basis for these differences we have developed a generally
applicable fluorescence-based technique for real-time monitoring
of the activation switch of GPCRs in living cells. We used such direct
measurements to investigate the activation of the alpha(2A)-adrenergic
receptor (alpha(2A)AR; neurotransmitter) and the parathyroid hormone
receptor (PTHR; hormone) and observed much faster kinetics than expected:
approximately 40 ms for the alpha(2A)AR and approximately 1 s for
the PTHR. The different switch times are in agreement with the different
receptors' biological functions. Agonists and antagonists could rapidly
switch the receptors on or off, whereas a partial agonist caused
only a partial signal. This approach allows the comparison of agonist
and partial agonist intrinsic activities at the receptor level and
provides evidence for millisecond activation times of GPCRs.},
added-at = {2010-12-14T18:12:02.000+0100},
author = {Vilardaga, J. P. and Bunemann, M. and Krasel, C. and Castro, M. and Lohse, M. J.},
biburl = {https://www.bibsonomy.org/bibtex/24a4c24f0a7360e013477b3a37f34d533/pharmawuerz},
endnotereftype = {Journal Article},
interhash = {0c0b71c3f46efe16302afdfc3d004471},
intrahash = {4a4c24f0a7360e013477b3a37f34d533},
issn = {1087-0156 (Print) 1087-0156 (Linking)},
journal = {Nat Biotechnol},
keywords = {& 1/agonists/antagonists Binding Cell Clonidine/pharmacology Culture Fluorescence/*methods G-Protein-Coupled/agonists/antagonists GTP-Binding Hormone, Hormone/pharmacology Humans Kidney/chemistry/drug Line Parathyroid Protein Proteins/*chemistry/*metabolism Signal Spectrometry, Techniques/*methods Transduction/*physiology Type alpha-2/agonists/antagonists effects/metabolism inhibitors/*chemistry/*metabolism inhibitors/chemistry inhibitors/metabolism Receptor Adrenergic},
month = Jul,
note = {Vilardaga, Jean-Pierre Bunemann, Moritz Krasel, Cornelius Castro,
Marian Lohse, Martin J Comparative Study Research Support, Non-U.S.
Gov't United States Nature biotechnology Nat Biotechnol. 2003 Jul;21(7):807-12.
Epub 2003 Jun 15.},
number = 7,
pages = {807-12},
shorttitle = {Measurement of the millisecond activation switch of G protein-coupled
receptors in living cells},
timestamp = {2010-12-14T18:22:43.000+0100},
title = {Measurement of the millisecond activation switch of G protein-coupled
receptors in living cells},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12808462},
volume = 21,
year = 2003
}