New Tool Lets Researchers Watch Drug Receptors Change Shape in Living Cells
Scientists have developed a technique to observe how drug receptors move and shift when medications bind to them—in real time, inside living cells. The advance could accelerate drug discovery by letting companies test thousands of compounds faster and identify which ones work best at the molecular level.
Originaltitel: Conformational GPCR BRET Sensors Based on Bioorthogonal Labeling of Noncanonical Amino Acids.
Here we describe the application of genetic code expansion and site-specific incorporation of noncanonical amino acids that serve as anchor points for fluorescent labeling to generate bioluminescence resonance energy transfer (BRET)-based conformational sensors. Using a receptor with an N-terminal NanoLuciferase (Nluc) and a fluorescently labeled noncanonical amino acid in the receptor's extracellular part allows to analyze receptor complex formation, dissociation, and conformational rearrangements over time and in living cells. These BRET sensors can be used to investigate ligand-induced intramolecular (cysteine-rich domain [CRD] dynamics), but also intermolecular (dimer dynamics) receptor rearrangements. With the design of BRET conformational sensors based on the minimally invasive bioorthogonal labeling procedure, we describe a method that can be used in a microtiter plate format and can be easily adopted to investigate ligand-induced dynamics in various membrane receptors.