Extended Data Fig. 12: MRGPRX2 interactions with Gi and Gq.
From: Structure, function and pharmacology of human itch receptor complexes
a, Effects of mutations in the MRGPRX2 along the Gi trimer interface on PAMP-12 induced Gαi–Gγ dissociation. Values are the mean ± SEM of three independent experiments for the wild type (WT) and mutants (n=3). Statistical differences between WT and mutations were determined by two-sided one-way ANOVA with Tukey test. *, P < 0.05; **, P < 0.01; ***, P < 0.001, n.s., no significant difference. (P < 0.0001, < 0.0001, 0.0022, 0.0018, < 0.0001, 0.0008, < 0.0001, 0.0395, 0.0009, 0.0009, 0.0003, 0.0006, 0.0010, 0.0178, 0.0018, 0.0003, 0.0003 from from top to bottom). b, RMSD of PAMP-12–MRGPRX2 (upper panel) and Gαi (lower panel) during 200 ns MD simulations. c,RMSD of PAMP-12–MRGPRX2 (upper panel) and Gαq (lower panel) during 200 ns MD simulations. d, PAMP-12–MRGPRX2–Gαq complex model after 200 ns MD simulation using M1R-Gq/G11 complex as the Gq template (PDB: 6OIL). e, Structural comparison of PAMP-12–MRGPRX2–Gαq stimulated model (orange) and PAMP-12–MRGPRX2–Gi complex (green). f, Effects of mutations in the MRGPRX2 along the Gαq trimer interface on PAMP-12 induced Gαq–Gγ protein dissociation. Values are the mean ± SEM of three independent experiments for the wild type (WT) and mutants (n=3). Statistical differences between WT and mutations were determined by two-sided one-way ANOVA with Tukey test. *, P < 0.05; **, P < 0.01; ***, P < 0.001, n.s., no significant difference. (P < 0.0001, 0.0016, < 0.0001, < 0.0001, 0.9226, 0.8226, 0.8815, < 0.0001, < 0.0001, < 0.0001, < 0.0001, < 0.0001, < 0.0001, < 0.0001, < 0.0001, < 0.0001, 0.0319 from top to bottom). g-i, Because the OD1 and OD2 in the carboxylate group of aspartic acid (Asp), or OE1 and OE2 in the carboxylate group of glutamic acid (Glu), or the NH1 and NH2 in guanidinium group of arginine (Arg) are equal atoms respectively, we named the ODc:NHC pair as closer pair for the contacting Asp: Arg, OEC:NHC pair as closer pair for the contacting Glu: Arg; and the remote ODR atom, OER atom, or NHR atom was named in Asp, Glu or Arg respectively g, The average distances between NHC in R140 of MRGPRX2 and oxygen atom of N198 of Gαq in MRGPRX2–Gαq complex or nitrogen atom of the guanidium group of R140 (NE) of MRGPRX2 and ODC of D193 of Gαi1 in MRGPRX2–Gi1 complex calculated from triplicate 200 ns MD simulations. h, The average distances between NHC in R143 of MRGPRX2 and NHC in R24 of Gαq in MRGPRX2–Gαq complex or NHC in R143 of MRGPRX2 and OEC in E28 of Gαi1 in MRGPRX2–Gαi1 complex calculated from triplicate 200 ns MD simulations. i, The average distances between NHC in R214 of MRGPRX2 and carbon atom of the methyl group I323(CD) in MRGPRX2–Gq complex or NHC in R214 of MRGPRX2 and OEC in E318 of Gαi1 in MRGPRX2–Gαi1 complex calculated from triplicate 200 ns MD simulations. j: The effects of selected SNPs on ligand-induced MRGPRX2 activation. SNPs located in the ligand or Gi protein interface of MRGPRX2 are summarized. The Gαi/Gαq–Gγ dissociation assays were measured in the current study, whereas calcium mobilization and mast cell degranulation were obtained from previous studies20 .