Figure 3.

Interaction sites of Rg5 with P2RY₁₂. (A) P2RY₁₂ and Rg5 complex structure, and binding pocket with Rg5. (B) Two-dimensional interaction map of interactions between Rg5 and P2RY₁₂. (C) Energy decomposition of amino acid residues in P2RY₁₂ and Rg5 based on MM/PBSA free energy calculation. (D) Structures of P2RY₁₂ mutation variants with Rg5 bound were shown in the upper panel, and Western blot analysis in the lower panel displayed the protein expression and phosphorylation levels of AKT (n = 4) and ERK (n = 4). All the data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. 2MesADP group. (E) Hypothetical binding modes of Rg5 to P2RY₁₂ mutation variants obtained by molecular docking. The binding conformation of Rg5 in each P2RY₁₂ mutant were colored as follows: white, original; slate, E188A; orange, E188A and R265A; magenta, R165A; pink, D266A; cyan, E188A and D266A; yellow, R265A and D266A; green, E188A and R265A and D266A. (F) A view of the P2RY12 mutant/Rg5 complex.