Aptamer-assisted two-point immobilized agonist-bound angiotensin II type 1 receptor for a second-site modulator discovery

2023 Julius Axelrod Symposium: Plant-Derived Molecules Acting on G Protein-Coupled Receptors

“…Monelin, a protein isolated from Dioscoreophyllum cumminsii, was identified as the first naturally-derived allosteric agonist of metabotropic glutamate receptor 5 binding to its large extracellular domain (Chen et al, 2020). An allosteric effect was also reported on the AT1 receptor for Ginsenoside Rg1, an active component extracted from Salviae Miltiorrhizae Radix et Rhizoma, which is suggested to bind to an allosteric site in the extracellular loop 2 (Liu et al, 2022). Similarly, Muratspahic et al, showed that T20K, a plant-derived cyclotide in clinical development for the treatment of multiple sclerosis, is a positive allosteric modulator of the κ-OR by using binding and functional assays (Muratspahić et al, 2021b).…”

“…Other binding techniques such as the label-free surface plasmon resonance (SPR) technique have been employed recently to confirm hits from virtual screens as shown for the bisbenzylisoquinoline alkaloid neferine and orexin receptors (He et al, 2023). Recent progress in the expression and purification of recombinant GPCRs helped to determine the binding properties or to purify plant-derived molecules by affinity chromatography on immobilized GPCRs (AT1, ETA, M3 receptors) (Fan et al, 2023;Liang et al, 2020;Liang et al, 2021;Liu et al, 2022;Shayiranbieke et al, 2022;Zhang et al, 2023).…”

“…This technique has been successfully applied to the β1-AR (Shayiranbieke et al, 2022), the ETA receptor (Ji et al, 2023) and the M3 muscarinic receptor (Fan et al, 2023) to screen candidate ligands and to identify receptor binding molecules in herbal extracts. An alternative, two-point immobilization protocol has been also described for the AT1 receptor (Liu et al, 2022). In this study, the authors utilized two single-stranded DNA strands to immobilize a selective AT1 aptamer and the angiotensin II (3-8) peptide separately to the microsphere surface.…”