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Updated: May 21, 2025

A Kinetic Fluorescence-based Ca2+ Mobilization Assay to Identify G Protein-coupled Receptor Agonists, Antagonists, and Allosteric Modulators
Published on: February 20, 2018
Activation dynamics traced through a G protein-coupled receptor by 81 1H-15N NMR probes
Feng-Jie Wu1, Pascal S Rieder2, Layara Akemi Abiko1
1Biozentrum, University of Basel, Basel, Switzerland.
Abstract:
The regulation of G protein-coupled receptor signaling by different orthosteric ligands is thought to occur through shifts in dynamically interconverting, conformational distributions. Such changes in dynamical distributions have been detected so far only by very sparse, often non-native experimental probes at low resolution. Using a recently developed paramagnetic nuclear magnetic resonance (NMR) method, we could assign and follow 81 1H-15N NMR correlations in the β1-adrenergic receptor β1AR at ambient conditions in response to various orthosteric ligands in the absence or presence of a G protein-mimicking nanobody. The comparison reveals the dynamics and mechanism of the central, highly conserved xWIPF3 motif, contiguous regions of rigid and loose conformational coupling separated by conserved prolines during signal transmission, and the plasticity of the intracellular face in response to transducer binding.
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