Luteinizing hormone and chorionic gonadotropin are glycoprotein hormones that are related to follicle-stimulating hormone and thyroid-stimulating hormone1,2. Luteinizing hormone and chorionic gonadotropin are essential to human reproduction and are important therapeutic drugs3,4,5,6. They activate the same G-protein-coupled receptor, luteinizing hormone–choriogonadotropin receptor (LHCGR), by binding to the large extracellular domain3. Here we report four cryo-electron microscopy structures of LHCGR: two structures of the wild-type receptor in the inactive and active states; and two structures of the constitutively active mutated receptor. The active structures are bound to chorionic gonadotropin and the stimulatory G protein (Gs), and one of the structures also contains Org43553, an allosteric agonist7. The structures reveal a distinct ‘push-and-pull’ mechanism of receptor activation, in which the extracellular domain is pushed by the bound hormone and pulled by the extended hinge loop next to the transmembrane domain. A highly conserved 10-residue fragment (P10) from the hinge C-terminal loop at the interface between the extracellular domain and the transmembrane domain functions as a tethered agonist to induce conformational changes in the transmembrane domain and G-protein coupling. Org43553 binds to a pocket of the transmembrane domain and interacts directly with P10, which further stabilizes the active conformation. Together, these structures provide a common model for understanding the signalling of glycoprotein hormone receptors and a basis for drug discovery for endocrine diseases.
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The density maps and structure coordinates have been deposited in the Electron Microscopy Database (EMDB) and the PDB with accession numbers as follows: EMD-31596, PDB 7FIG for the CG–LHCGR(S277I)–Gs complex; EMD-31597 and 7FIH for the CG–Org43553–LHCGR(S277I)–Gs complex; EMD-31598 and 7FII for the CG–LHCGR(WT)–Gs complex; and EMD-31599 and 7FIJ for the LHCGR (inactive) structure.
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The cryo-EM data were collected at the Cryo-Electron Microscopy Research Center, Shanghai Institute of Materia Medica (SIMM) and Shuimu BioSciences. We thank the staff at the SIMM Cryo-Electron Microscopy Research Center and Shuimu BioSciences for their technical support. This work was partially supported by Ministry of Science and Technology (China) grants (2018YFA0507002 to H.E.X. and 2019YFA0508800 to Y.Z.); the Shanghai Municipal Science and Technology Major Project (2019SHZDZX02 to H.E.X.); the CAS Strategic Priority Research Program (XDB08020303 to H.E.X.); the National Natural Science Foundation of China (31770796 to Y.J. and 81922071 to Y.Z.); the National Science and Technology Major Project ‘Key New Drug Creation and Manufacturing Program’ (2018ZX09711002 to Y.J. and H.J.); Wellcome Trust grant 209407/Z/17/Z; the National Key R&D Program of China (2020YFC0861000 to S.Z.); the CAMS Innovation Fund for Medical Sciences (2020-I2M-CoV19-001 to S.Z.); the Tsinghua University–Peking University Center for Life Sciences (045-160321001 to S.Z.); the Science and Technology Commission of Shanghai Municipal (20431900100 to H.J.); the Jack Ma Foundation (2020-CMKYGG-05 to H.J.); Shanghai Sailing Program (19YF1457600 to Q.L.); the Zhejiang Province Science Fund for Distinguished Young Scholars (LR19H310001 to Y.Z.); the MOE Frontier Science Center for Brain Science & Brain-Machine Integration, Zhejiang University (to Y.Z.); and The Youth Innovation Promotion Association of Chinese Academy of Science (2018319 to X.C).
The authors declare no competing interests.
Peer review information Nature thanks Torsten Schöneberg and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Peer reviewer reports are available.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Extended data figures and tables
a, Size-exclusion chromatography elution profile and SDS-PAGE of the CG–LHCGR–Gs complex. Red star indicates the monomer peak of the complex. For gel source data, see Supplementary Fig. 1 (Experiments were repeated three times with similar results). b, Cryo-EM micrograph, reference-free 2D class averages, and flowchart of cryo-EM data analysis of the CG–LHCGR–Gs complex. c, Cryo-EM map of the CG–LHCGR–Gs complex coloured by local resolutions from 2.0 Å (blue) to 5.0 Å (red). The ‘gold-standard’ Fourier shell correlation (FSC) curve indicates that the overall resolution of the electron density map of the CG–LHCGR–Gs complex is 3.9 Å.
Extended Data Fig. 2 Cryo-EM image and single-particle reconstruction of the Org43553–CG–LHCGR–Gs complex.
a, Size-exclusion chromatography elution profile and SDS–PAGE of the Org43553–CG–LHCGR–Gs complex. Red star indicates the monomer peak of the two complex. For gel source data, see Supplementary Fig. 1 (Experiments were repeated three times with similar results). b, c, Cryo-EM micrograph, reference-free 2D class averages, and flowchart of cryo-EM data analysis of the Org43553–CG–LHCGR–Gs complex. Org43553–CG–LHCGR–Gs complex coloured by local resolutions from 2.0 Å (blue) to 5.0 Å (red).The ‘gold-standard’ Fourier shell correlation (FSC) curves indicate that the overall resolution of the Org43553–CG–LHCGR–Gs complex is 3.18 Å (Focused refinement) and 3.3 Å (Global refinement), respectively. d, Cryo-EM density map and model of the Org43553–CG–LHCGR–Gs complex. The regions of the cryo-EM density map with all transmembrane helices, H8 and hinge C terminus are shown.
Extended Data Fig. 3 Cryo-EM images and single-particle reconstruction of the Org43553–CG–LHCGR(WT)–Gs complex and inactive LHCGR, and electron microscopy maps for the inactive LHCGR structure.
a, b, Size-exclusion chromatography elution profiles and SDS-PAGEs of the Org43553–CG–LHCGR(WT)–Gs complex (left panel) and inactive LHCGR (right panel). Red stars indicate the monomer peaks of the two proteins. For gel source data, see Supplementary Fig. 1 (Experiments were repeated three times with similar results). c, d, Cryo-EM micrograph, reference-free 2D class averages, and flowchart of cryo-EM data analysis of the Org43553–CG–LHCGR(WT)–Gs complex (c) and inactive LHCGR (d). e, f, Cryo-EM maps of the Org43553–CG–LHCGR(WT)–Gs complex (e) and inactive LHCGR (f) coloured by local resolutions from 2.0 Å (blue) to 5.0 Å (red). The “Gold-standard” Fourier shell correlation (FSC) curves indicate that the overall resolution of the electron density map of the Org43553–CG–LHCGR(WT)–Gs complex is 4.3 Å (e) and inactive LHCGR is 3.8 Å (f). g, Cryo-EM density map and model of the inactive LHCGR. The regions of the cryo-EM density map with all transmembrane helices and H8 are shown.
a, Ribbon presentation of CGα and CGβ subunits stabilized by cysteine-knots. Disulfide bonds are shown as yellow sticks. b, Ribbon presentation of CGα and CGβ subunits. The C terminus of the CGβ subunit, known as ‘seat belt’ highlighted in red, circulates the CGα subunit. c, A ‘hand-clasp’ binding fashion of CG to LHCGR from different views. CGα and CGβ subunits are shown in a surface presentation, and LHCGR is displayed as a ribbon. d, Structural comparison of free human CG (PDB code: 1HCN, green–yellow) with receptor-bound CG (light sea green), the conformational changes in four segments are highlighted in red rectangle. e, Structural comparison of seat belt between CG–LHCGR and FSH–FSHR (PDB code: 1XWD). The residue P107 in CGβ and P101 in FSHβ are shown as sticks. f, Conformational comparison of TM6 of inactive LHCGR with inactive rhodopsin (PDB code:1L9H, left panel) and inactive β2AR (PDB code: 5JQH, right panel). g, Electrostatic potential surface of CG. The positively charged pockets that interact with the hinge loop are highlighted in red circles. h, Concentration–response curves for point mutants at S277. Data from three independent experiments are presented as the mean ± s.e.m. i, The putative interaction between S277 in the hinge helix and N351 in the P10 region based on the model of the Org43553–CG–LHCGR–Gs complex.
a, Sequence alignment of human FSHR, LHR and TSHR in the region of the hormone-binding domain. Residues only interact with CGβ (light sea green), CGα (pink), and both CGα and CGβ subunits (light blue) are highlighted. Residues that determine LHCGR specificity were labeled with asterisks. b, Sequence alignment of human CG, LH, TSH and FSH β-subunit. CGβ residues interacted with LHCGR are highlighted in light sea green. c, The α-subunit sequence of glycoprotein hormones. CGα residues interacted with LHCGR are highlighted in pink.
a, Representative snapshots from the CG-bound receptor simulation and inactive receptor simulation, respectively. The receptor is shown as cartoon, the CG is shown as surface, the phosphate groups of the lipid membrane are shown as spheres, three ECD residues N107, D157 and E206 are shown as sticks. b, Changes of ECD orientations observed in the different simulations as a function of time. Tilting angle of the ECD with respect to the z-axis (membrane normal) and the minimal distance from an ECD residue (D157, N107, or E206) to the membrane. c, Structural alignment of the CG–LHCGR complex onto the FSH–FSHR-ECD dimer (PDB code: 1WXD, left panel) shows that the two TMD from each complex is pointed in the opposite direction and cannot be on the same membrane layer. d, Structural alignment of the CG–LHCGR complex onto the FSH–FSHR-ECD trimer (PDB code: 4AY9, right panel) shows that the TMD from the CG–LHCGR complex will clash with the neighbouring ECD in the trimeric arrangement. The trimeric FSH–FSHR ECD was shown in surface presentation, with each monomeric complex in a single colour (red, yellow, green). The CG–LHCGR is shown as a ribbon and is aligned onto the yellow ECD complex but is clashed with the red ECD complex.
a, Two views of the Org43553-binding pocket in the representative structures of molecular dynamics Simulation 1 and 2. Org43553 is shown as sticks, and LHCGR is depicted by surface. b, The interactions between I585, S604 and Org43553 in the cryo-EM structure and the representative structures of simulation 1 and simulation 2. Org43553 is shown as sticks, and LHCGR is shown as cartoon. c, The time-course curve of the shortest distance between the heavy atoms of I585, S604 and Org43553 during simulation 1 and 2. d, The distribution of the root mean square deviation (RMSD) from the cryo-EM ligand pose in simulations of Org43553-bound LHCGR system. e, Comparison of the Org43553-binding pocket with other agonist-binding pockets of class A GPCRs. β2AR (PDB code: 3SN6); β1AR (PDB code: 2Y03); A2aR (PDB code: 3QAK); 5HT1B (PDB code: 4IAR); Org43553, tomato; β2AR-agonist, yellow; β1AR-agonist, cyan; A2aR-agonist, lime green; 5HT1B-agonist, magenta.
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Duan, J., Xu, P., Cheng, X. et al. Structures of full-length glycoprotein hormone receptor signalling complexes. Nature 598, 688–692 (2021). https://doi.org/10.1038/s41586-021-03924-2
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