Fig. 2. Analysis of the allosteric binding site of xanomeline.

a Xanomeline is bound in both the orthosteric and allosteric binding sites of the M₄ mAChR (M4R). b Cryo-EM density (contour level 0.026) for xanomeline in the allosteric binding site. c Xanomeline in the common ‘ECV’ mAChR allosteric binding site with allosteric site residues shown as sticks in green. d Comparison of the xanomeline allosteric binding site to LY2033298 and VU0467154 bound to the allosteric binding site of the M₄ mAChR (PDB: 7TRP and 7TRQ). The allosteric binding site residues of the xanomeline bound M₄ mAChR are shown as green sticks whereas the allosteric binding site residues of the LY2033298 and VU0467154 bound M₄ mAChR are shown in blue and grey sticks, respectively. e Comparison of the xanomeline allosteric binding site to LY2119620 bound to the allosteric binding site of the M₂ mAChR (M2R, PDB: 6OIK). The allosteric binding site residues of the M₄ mAChR are shown as green sticks whereas the allosteric binding site residues of the M₂ mAChR are shown in light blue sticks. f Overlay of M₄ mAChR bound xanomeline, LY2033298, VU0467154 and M₂ mAChR bound LY2119620.