%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 }