title: An optical dynamic mass redistribution assay reveals biased signaling of dualsteric GPCR activators.
abstract: Increasing attention is paid in basic science and in drug discovery to pathway selective intracellular signaling as a novel approach to achieve precise control of cell function via G protein-coupled receptors (GPCRs). With respect to signaling, GPCRs are often promiscuous in that more than one intracellular biochemical pathway is activated upon receptor stimulation by the endogenous transmitter or by exogenous drugs. We studied signaling by a novel class of GPCR activators that were designed to bind simultaneously to the orthosteric transmitter-binding site and the allosteric site of muscarinic acetylcholine receptors. An optical biosensor technique was applied to measure activation-induced dynamic mass redistribution (DMR) in CHO cells stably expressing the muscarinic receptor subtype of interest. The use of tools to modulate signaling and measuring G protein activation directly proved that DMR is a valid and comfortable approach to gain real-time insight into intracellular signaling pathway activation and to identify signaling pathway-selective drugs.
authors: Pharmacology and Toxicology Section, Institute of Pharmacy, Rheinische Friedrich-Wilhelms-University, Bonn, Germany.
authors: Anna
authors: A
authors: Kebig
authors: Evi
authors: E
authors: Kostenis
authors: Klaus
authors: K
authors: Mohr
authors: Marion
authors: M
authors: Mohr-Andrä
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D015374:Biosensing Techniques; D016466:CHO Cells; D002217:Carbachol / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D018679:Cholinergic Agonists / Q000494:pharmacology; D006224:Cricetinae; D003412:Cricetulus; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D018721:Muscarinic Agonists / Q000494:pharmacology; D018727:Muscarinic Antagonists / Q000494:pharmacology; D010095:Oxotremorine / Q000031:analogs & derivatives / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology
chemical_list: D018679:Cholinergic Agonists; D018721:Muscarinic Agonists; D018727:Muscarinic Antagonists; D043562:Receptors, G-Protein-Coupled; D010095:Oxotremorine; C042743:oxotremorine M; D001285:Atropine; D002217:Carbachol; D000109:Acetylcholine