Extended Data Table 1.

Cryo-EM data collection, refinement and validation statistics

	NTS-NTSR1-Gi-cND canonical state, AHD and nanodisc mask out (EMD-23099, PDB 7L0P)	NTS-NTSR1-Gi-cND canonical state, overall (EMD-23100, PDB 7L0Q)	NTS-NTSR1-Gi-cND noncanonical state, AHD and nanodisc mask out (EMD-23101, PDB 7L0R)	NTS-NTSR1-Gi-cND noncanonical state, overall (EMD-23102, PDB 7L0S)
Data collection and processing
Magnification	105,000	105,000	105,000	105,000
Voltage (kV)	300	300	300	300
Electron exposure (e-/Å2)	57	57	57	57
Defocus range (μm)	−1.2 to −2.5	−1.2 to −2.5	−1.2 to −2.5	−1.2 to −2.5
Pixel size (Å)	0.825	0.825	0.825	0.825
Symmetry imposed	C1	C1	C1	C1
Initial particle images (no.)	4,367,542	4,367,542	4,367,542	4,367,542
Final particle images (no.)	575,791	575,791	324,002	324,002
Map resolution (Å)	4.1	4.3	4.2	4.5
FSC threshold	0.143	0.143	0.143	0.143
Refinement
Initial model used (PDB code)	4BUO, 1GP2, 6OY9
Map sharpening B factor (Å2)	−245	−222	−204	−228
Model composition
Non-hydrogen atoms	6954	7892	6968	7906
Protein residues	882 (6896 atoms)	996 (7834 atoms)	881 (6910 atoms)	995 (7848 atoms)
Ligands	1 (6 residues, 58 atoms)	1 (6 residues, 58 atoms)	1 (6 residues, 58 atoms)	1 (6 residues, 58 atoms)
B factors (Å2)
Protein	73.6	69.75	73.63	69.79
Ligand	66.76	66.76	66.76	66.76
R.m.s. deviations
Bond lengths (Å)	0.006	0.007	0.007	0.008
Bond angles (°)	1.051	1.286	1.368	1.508
Validation
MolProbity score	1.9	1.84	1.96	2
Clashscore	8.36	7.82	9.79	9.27
Poor rotamers (%)	0.53	0.7	1.05	1.39
Ramachandran plot
Favored (%)	93.1	93.69	93.44	93.99
Allowed (%)	6.9	6.31	6.56	6.01
Disallowed (%)	0	0	0	0