Fig. 1: Structure of constitutively active FZD7 in complex with heterotrimeric mGs. | Cell Research

Fig. 1: Structure of constitutively active FZD7 in complex with heterotrimeric mGs.

From: Cryo-EM structure of constitutively active human Frizzled 7 in complex with heterotrimeric Gs

Fig. 1

a Normalized BRET0 values of ΔFZD1–10 HEK293 cells transiently co-transfected with the Gs BRET sensor along with either negative control (mock), the β2-adrenoceptor (β2AR) or FZD7. Data are represented as the means ± SEM of raw BRET0 that were obtained from simple linear regression of five independent experiments measured in quadruplicates shown in Supplementary information, Fig. S1A and normalized to the negative control. *P < 0.05; **P < 0.01 (one-way ANOVA followed by Sidak’s multiple comparison). b An annotated 2D class average of FZD7–mGs–Nb35 complex. c Overall density map and atomic model of FZD7–mGs–Nb35 complex (CRD was omitted due to linker flexibility). FZD7, blue; mGαs, orange; Gβ, green; Gγ, yellow; Nb35, gray. d Insertion of the α5-helix (mGαs, orange) into FZD7 helical bundle represented as surface (ICL1, blue; ICL2, pink; ICL3, yellow; TM7/H8, green). e Schematics of interactions between FZD7 and α5-helix. Hydrogen bonds are shown as red dashed lines. The red circle represents the hydrophobic interaction network. Yellow shades indicate residues that reside in TM5/6/ICL3; pink, TM3/4/ICL2; blue, TM1/2/ICL1; green, TM7/H8. f Superposition of FZD7 (blue) and FZD4 (yellow) structures, viewed from the intracellular side (bottom view). g Superposition of the active FZD7 structure (blue) with the inactive FZD4 (PDB: 6BD4, yellow), inactive FZD5 (PDB: 6WW2, light pink), active SMO (PDB: 6OT0, gray) and inactive SMO (PDB: 5V57, green) structures. h Comparison of the cytoplasmic portion of TM6 (from K6.28 to P6.43) in FZD7, FZD4, active SMO and inactive SMO structures. i R6.32, F6.36, W7.55 network in FZD7, FZD4 and active SMO structures. Blue dashed lines indicate the distance of F6.36–W7.55 and F6.36–R6.32 in FZD7. Gray dashed lines indicate the distance of R6.32–W7.55 and W7.55–F6.36 in active SMO structure. j Normalized BRET0 values of ΔFZD1-10 HEK293 cells transiently co-transfected with rGFP-CAAX and Gαs-67-RlucII, along with either negative control (mock), wild-type FZD7, ΔCRD-FZD7 or the indicated FZD7 mutants. Data are represented as the means ± SEM of raw BRET0 that were obtained from simple linear regression of four independent experiments measured in quadruplicates shown in Supplementary information, Fig. S11b and normalized to the negative control. **P < 0.01; ***P < 0.001 (one-way ANOVA followed by Tukey’s multiple comparison). k Normalized FRET0 values of ΔFZD1–10 HEK293 cells transiently co-transfected with the FRET-based cAMP biosensor along with either negative control (mock), wild-type FZD7, ΔCRD-FZD7 or the indicated FZD7 mutants. Data are represented as the means ± SEM of raw FRET0 that were obtained from simple linear regression of five independent experiments measured in quadruplicates shown in Supplementary information, Fig. S11c and normalized to the negative control. **P < 0.01; ****P < 0.0001 (one-way ANOVA followed by Sidak’s multiple comparison).

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