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. Author manuscript; available in PMC: 2010 Mar 1.
Published in final edited form as: Nat Chem Biol. 2009 Aug 2;5(9):688–695. doi: 10.1038/nchembio.199

Figure 1. Functional complementation of two “non-functional receptors”.

Figure 1

We used an aequorin assay that couples Gq (or Gqi5) activation to a luminescence readout. (a) The agonist quinpirole did not lead to D2R-induced Gq activation. (b) D2R when coexpressed with free Gqi5 or (c) D2R fused with Gqi5 via a linker (D2-linker-Gqi5) led to quinpirole-induced luminescence. (c) A nonfunctional Gα deficient fusion construct, D2-linker-Gqi5G208A failed to produce luminescence. (d) Free Gqi5 rescued the function of D2-linker-Gqi5G208A. (e) Free Gqi5 failed to rescue the function of non-linker D2R-Gqi5, which unlike D2R-linker-Gqi5, did not signal when expressed alone. (f) Coexpressing D2R with D2R-Gqi5 (12 hour tetracycline induction) restored signaling, despite the inability of either construct to signal in this assay when expressed alone. Activation data represent luminescence relative to that seen with 0.1% triton treatment. The mean±SEM of at least 3 experiments, each conducted in triplicate, are shown. The symbols used in (Fig. 1–Fig. 4 and Fig. 6) are explained in detail in (Supplementary Fig. 2 online).