Journal: Nature Communications

Article Title: Structural and dynamic studies uncover a distinct allosteric modulatory site at the µ-opioid receptor

doi: 10.1038/s41467-026-72633-z

Figure Lengend Snippet: A Overview of BMS-986187/DAMGO/MOR-Gi1 complex and receptor-only structures built based on cryo-EM density of the complex. Zoom-in shows BMS-986187 (orange) and DAMGO (magenta) bound at the receptor. MOR, Gαi1, Gβ1, and Gγ2, are shown in palecyan, wheat, green, and purple, respectively. Ligands are shown as stick models with transparent spheres. B Allosteric effect of BMS-986187 on DAMGO signaling in cAMP accumulation assays, Gi1 BRET assays, and β-arrestin2 recruitment assays. Data represent mean ± SEM of three independent experiments ( n = 3) performed in triplicate, and have been normalized to the DAMGO + vehicle response. C Intrinsic efficacy of BMS-986187, DAMGO, and vehicle (DMSO) at MOR as determined by cellular cAMP accumulation assay. Data represent mean ± SEM of four independent experiments ( n = 4) performed in triplicate, and have been normalized to the DAMGO response. Source data are provided as a file.

Article Snippet: After 24 h, media was removed and 0.4 mg/ml D-luciferin (Gold Biotechnology) was loaded into cells in 15 μl of Hank’s balanced salt solution (HBSS) supplemented with 20 mM HEPES pH 7.4, 0.1% BSA (w/v) and 0.01% ascorbic acid (w/v), and cells were incubated for 1 h. DAMGO (Tocris Bioscience) and BMS-986187 (MedChemExpress) were dissolved in DMSO to prepare 10 mM stock solutions.

Techniques: Cryo-EM Sample Prep

Journal: Nature Communications

Article Title: Structural and dynamic studies uncover a distinct allosteric modulatory site at the µ-opioid receptor

doi: 10.1038/s41467-026-72633-z

Figure Lengend Snippet: A Left, structure of the BMS-986187-bound DAMGO/MOR-Gi1 signaling complex (PDB: 9PU5). Middle, BMS-986122-bound DAMGO/MOR-Gi3 signaling complex (PDB: 8K9L). Right, compound 368-bound naloxone/MOR-Nb6 inactive-state complex (PDB: 9BJK). Zoom-ins represent extracellular views of the orthosteric binding pockets. PAMs, orthosteric ligands, MOR, Gαi1, Gβ1, and Gγ2, are shown in orange, magenta, palecyan, wheat, green, and purple, respectively. Ligands are shown as stick models with transparent spheres. B Chemical structures of select positive allosteric modulators potentiating MOR activity.

Article Snippet: After 24 h, media was removed and 0.4 mg/ml D-luciferin (Gold Biotechnology) was loaded into cells in 15 μl of Hank’s balanced salt solution (HBSS) supplemented with 20 mM HEPES pH 7.4, 0.1% BSA (w/v) and 0.01% ascorbic acid (w/v), and cells were incubated for 1 h. DAMGO (Tocris Bioscience) and BMS-986187 (MedChemExpress) were dissolved in DMSO to prepare 10 mM stock solutions.

Techniques: Binding Assay, Activity Assay

Journal: Nature Communications

Article Title: Structural and dynamic studies uncover a distinct allosteric modulatory site at the µ-opioid receptor

doi: 10.1038/s41467-026-72633-z

Figure Lengend Snippet: A Interactions between BMS-986187 (orange), MOR (palecyan), and cholesterol (CLR, green). The left panel highlights interface residues identified in the cryo-EM structure. The middle panel shows the frequency of prevalent interactions (interaction frequency >0.35) observed during MD simulations. Blue shading over a bar indicates that the interaction is also observed in the cryo-EM structure. Interactions involving individual ligand atoms were grouped by molecular fragment, as illustrated in the right panel. B Interactions between MOR and a cholesterol molecule identified in the cryo-EM structure. C Interactions between DAMGO (magenta), bound in the orthosteric site, and MOR. Bar heights reflect the mean interaction frequency across all simulation frames which are based on 20 replicates of 450 ns each. Error bars represent the standard deviation (SD) computed over 25 ns time blocks (i.e., 18 per replicate) across all replicates. The initial and final coordinates of each MD simulation are available at https://github.com/filizolalab/bms187-init-end-md-structures .

Article Snippet: After 24 h, media was removed and 0.4 mg/ml D-luciferin (Gold Biotechnology) was loaded into cells in 15 μl of Hank’s balanced salt solution (HBSS) supplemented with 20 mM HEPES pH 7.4, 0.1% BSA (w/v) and 0.01% ascorbic acid (w/v), and cells were incubated for 1 h. DAMGO (Tocris Bioscience) and BMS-986187 (MedChemExpress) were dissolved in DMSO to prepare 10 mM stock solutions.

Techniques: Cryo-EM Sample Prep, Standard Deviation

Journal: Nature Communications

Article Title: Structural and dynamic studies uncover a distinct allosteric modulatory site at the µ-opioid receptor

doi: 10.1038/s41467-026-72633-z

Figure Lengend Snippet: A Binding site of BMS-986187 (orange), with MOR residues shown in cyan that were tested in cAMP accumulation assays. DAMGO concentration response data in the presence of vehicle (green), 3 μM BMS-986187 (red), and 10 μM BMS-986187 (blue) at wildtype and mutant MOR. Data represent mean ± SEM of three independent experiments ( n = 3) performed in triplicate, and have been normalized to the DAMGO + vehicle response. B Bar graphs showing the effect of mutations on DAMGO potency, expressed as potency difference between mutant and wildtype MOR (ΔpEC50 = pEC50 Mut – pEC50 WT ). Larger numbers indicate that DAMGO exhibits increased potency at mutant construct. C Bar graph showing PAM-mediated increases in DAMGO potency, expressed as potency difference at each mutant or wildtype between vehicle (DMSO) and 10 μM BMS-986187 (ΔpEC50 = pEC50 BMS-986187 – pEC50 DMSO ). Larger numbers indicate that BMS-986187 has an increased effect on DAMGO potency at the examined construct. D Bar graph showing intrinsic efficacy of BMS-986187 expressed as the difference in basal activity between vehicle (DMSO) and 10 μM BMS-986187 in DAMGO concentration response experiments. Larger numbers indicate a larger intrinsic efficacy of BMS-986187, and are shown as a fraction of the maximal efficacy observed in the system (% Emax). Data represent mean ± SEM calculated from three independent experiments ( n = 3), and graphs for B – D were obtained by performing a multiple t test between mean values from individual experiments. Source data are provided as a file.

Article Snippet: After 24 h, media was removed and 0.4 mg/ml D-luciferin (Gold Biotechnology) was loaded into cells in 15 μl of Hank’s balanced salt solution (HBSS) supplemented with 20 mM HEPES pH 7.4, 0.1% BSA (w/v) and 0.01% ascorbic acid (w/v), and cells were incubated for 1 h. DAMGO (Tocris Bioscience) and BMS-986187 (MedChemExpress) were dissolved in DMSO to prepare 10 mM stock solutions.

Techniques: Binding Assay, Concentration Assay, Mutagenesis, Construct, Activity Assay

Journal: Nature Communications

Article Title: Structural and dynamic studies uncover a distinct allosteric modulatory site at the µ-opioid receptor

doi: 10.1038/s41467-026-72633-z

Figure Lengend Snippet: A Representative structures of inactive- (PDB: 4DKL), active-like (PDB: 5C1M), active- (PDB: 6DDF) and BMS-986187-bound active-state (PDB: 9PU5) of MOR, with distinct rotamer configurations of Y151 and W194. B Representative structures highlight the binding of lipids and cholesterol near the BMS-986187 binding site, and show how BMS-986187 binding leads to a conformational shift in the cholesterol binding pose. Orthosteric ligand, BMS-986187, cholesterol/lipid, receptor and key residues Y151 and W194 are colored in yellow, orange, forest green, palecyan, and magenta respectively.

Article Snippet: After 24 h, media was removed and 0.4 mg/ml D-luciferin (Gold Biotechnology) was loaded into cells in 15 μl of Hank’s balanced salt solution (HBSS) supplemented with 20 mM HEPES pH 7.4, 0.1% BSA (w/v) and 0.01% ascorbic acid (w/v), and cells were incubated for 1 h. DAMGO (Tocris Bioscience) and BMS-986187 (MedChemExpress) were dissolved in DMSO to prepare 10 mM stock solutions.

Techniques: Binding Assay

Journal: Nature Communications

Article Title: Structural and dynamic studies uncover a distinct allosteric modulatory site at the µ-opioid receptor

doi: 10.1038/s41467-026-72633-z

Figure Lengend Snippet: A Total information flow (in bits) for three pathways: BMS-986187 to DAMGO, DAMGO to the G protein, and BMS-986187 to the G protein. Network diagrams illustrating the predominant allosteric communication pathways within MOR: from BMS-986187 to DAMGO ( B ), from DAMGO to the G protein ( C ), and from BMS-986187 to the G protein ( D ). The 20 residues with the highest contribution to information flow are labeled, including the nearby cholesterol molecule (labeled CLR). The top 10 contributing residues are highlighted in green and enclosed in boxes. Node size is proportional to the total information flow transmitted or received by each residue, while edge thickness indicates the magnitude of transfer entropy between residue pairs. Fractional information flow per residue (mean ± SEM, based on 20 samples of transfer entropy values) for the top 10 residues in each of the three pathways: BMS-986187 to DAMGO ( E ), DAMGO to G protein ( F ), and BMS-986187 to G protein ( G ). In F , flow values in the absence of BMS-986187 are shown in orange for comparison.

Article Snippet: After 24 h, media was removed and 0.4 mg/ml D-luciferin (Gold Biotechnology) was loaded into cells in 15 μl of Hank’s balanced salt solution (HBSS) supplemented with 20 mM HEPES pH 7.4, 0.1% BSA (w/v) and 0.01% ascorbic acid (w/v), and cells were incubated for 1 h. DAMGO (Tocris Bioscience) and BMS-986187 (MedChemExpress) were dissolved in DMSO to prepare 10 mM stock solutions.

Techniques: Labeling, Residue, Comparison