Roche
GTP-γ-S
10220647001
I use GTP-γ-S to measure the G-protein coupling affinity of serotonin 1A receptors because it allows me to assess the receptor's activity and function within the GPCR signaling pathway. GTP-γ-S is a non-hydrolyzable GTP analog that stabilizes the G-protein in its active state when bound to the receptor, enabling precise quantification of receptor-G-protein interactions. This technique will provide valuable insights into the coupling efficiency of serotonin 1A receptors with G-proteins, shedding light on their signaling capabilities and potential downstream effects.
Measurement of G-Protein coupling efficiency to GPCRs.
Cell membranes/native tissue expressing GPCR of interest
Use fresh dilutions from a concentrated stock before each experiment.
The effect of pertussis toxin treatment on G-protein coupling efficiency to serotonin 1A receptors was assessed using [3H]8-OH-DPAT binding in CHO-5-HT1A R cells. Monitoring the sensitivity of binding in the presence of increasing concentrations of GTP-γ-S revealed a characteristic reduction in specific binding for both control membranes (●) and membranes treated with pertussis toxin (▲).
https://doi.org/10.1016/j.bbamem.2020.183265
Robust assay for measuring G-protein coupling efficiency.
None.
GTP-γ-S is used to assess G-protein coupling affinity in GPCRs, providing valuable insights into their signaling activity and function.