Ligand-specific activation trajectories dictate GPCR signalling in cells
Nature • • business
Key Points:
- Researchers constructed M2 muscarinic receptor (M2R) biosensors by cloning human WT M2R into pcDNA3.0 with a cleavable signal peptide and fused fluorescent tags such as eGFP or HA-tags for expression and detection purposes.
- Site-directed mutagenesis introduced amber stop codons (TAG) at specific receptor positions to enable incorporation of noncanonical amino acids (ncAAs) using an orthogonal tRNA synthetase/tRNA system derived from Methanococcus barkeri, facilitating bioorthogonal labelling of receptors in live cells.
- HEK293T and HEK-AD cells were cultured and transfected with plasmids encoding M2R biosensors and the orthogonal translational machinery,
