Functional Stability of the Human Kappa Opioid Receptor Reconstituted in Nanodiscs Revealed by a Time-Resolved Scintillation Proximity Assay
Fig 5
Isolated KOR reconstituted in Nanodiscs activated Gi protein.
BODIPY FL GTPγS was used to confirm Gαi protein activation by KOR after isolation and reconstitution into Nanodiscs. The intrinsic activity of G proteins was measured in the presence of empty Nanodiscs (light grey, Empty ND). Compared to KOR/Nanodisc sample without agonist stimulation (KOR/ND, dark grey), the fluorescence from BODIPY FL GTPγS binding to Gαi increased by 56% when the KOR/Nanodisc sample was stimulated with 1 μM dynorphin A (1–17) (KOR/ND + Dyn A, dark grey). Error bars represent standard error of the mean between two independent experiments.