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. 2021 Apr 22;16(4):e0247743. doi: 10.1371/journal.pone.0247743

Fig 1. Addition of GDP disrupts GPCR-G protein complexes.

Fig 1

(A) Cartoon representation of the experimental design. Constitutively-active GPCRs fused to Renilla luciferase (Rluc8) form spontaneous active-state complexes with nucleotide-free G protein heterotrimers fused (via the Gβγ subunit) to the fluorescent protein Venus in the absence of activating ligands. Addition of GDP (100 μM) disrupts these complexes, decreasing BRET between GPCR-Rluc8 and Gαβγ-Venus. (B) Representative experiments of this type with GPR82 and GPR174. Traces represent the mean ± SD of 16 technical replicates from a single experiment, and each trace is normalized to the basal BRET observed for that particular G protein. GDP was injected where indicated by the horizontal bar.