Investigating the ligand agonism and antagonism at the D 2long receptor by dynamic mass redistribution

• Published in: Scientific reports Vol. 12; no. 1; p. 9637
• Main Authors: Forster, Lisa, Pockes, Steffen
• Format: Journal Article
• Language: English
• Published: England 10.06.2022
• Subjects: Animals; beta-Arrestins - metabolism; Cricetinae; Cricetulus; GTP-Binding Proteins - metabolism; Humans; Ligands; Receptors, G-Protein-Coupled - metabolism; Signal Transduction
• ISSN: 2045-2322
• DOI: 10.1038/s41598-022-14311-w

The signalling of the D receptor (D R), a G protein-coupled receptor (GPCR), is a complex process consisting of various components. For the screening of D R ligands, methods quantifying distinct second messengers such as cAMP or the interaction of the receptor with β-arrestin, are commonly employed. In contrast, a label-free biosensor technology like dynamic mass redistribution (DMR), where it is mostly unknown how the individual signalling pathways contribute to the DMR signal, provides a holistic readout of the complex cellular response. In this study, we report the successful application of the DMR technology to CHO-K1 cells stably expressing the human dopamine D receptor. In real-time kinetic experiments, studies of D R reference compounds yielded results for agonists and antagonists that were consistent with those obtained by conventional methods and also allowed a discrimination between partial and full agonists. Furthermore, investigations on the signalling pathway in CHO-K1 hD R cells identified the Gα protein as the main proximal trigger of the observed DMR response. The present study has shown that the DMR technology is a valuable method for the characterisation of putative new ligands and, due to its label-free nature, suggests its use for deorphanisation studies of GPCRs.