Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein Bias
<b>Background:</b> The four subtypes of G protein-coupled receptors (GPCRs) regulated by histamine play critical roles in various physiological and pathological processes, such as allergy, gastric acid secretion, cognitive and sleep disorders, and inflammation. Previous experimental stru...
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2025-02-01
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| author | Anqi Chen Chenxi Su Zisu Zhang Haitao Zhang |
| author_facet | Anqi Chen Chenxi Su Zisu Zhang Haitao Zhang |
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| description | <b>Background:</b> The four subtypes of G protein-coupled receptors (GPCRs) regulated by histamine play critical roles in various physiological and pathological processes, such as allergy, gastric acid secretion, cognitive and sleep disorders, and inflammation. Previous experimental structures of histamine receptors (HRs) with agonists and antagonists exhibited multiple conformations for the ligands and G protein binding. However, the structural basis for HR regulation and signaling remains elusive. <b>Methods:</b> We determined the cryo-electron microscopy (cryo-EM) structure of the H4R-histamine-Gi complex at 2.9 Å resolution, and predicted the models for all four HRs in the ligand-free apo and G protein subtype binding states using AlphaFold3 (AF3). <b>Results:</b> By comparing our H4R structure with the experimental HR structures and the computational AF3 models, we elucidated the distinct histamine binding modes and G protein interfaces, and proposed the essential roles of Y<sup>6.51</sup> and Q<sup>7.42</sup> in receptor activation and the intracellular loop 2 (ICL2) in G protein bias. <b>Conclusions:</b> Our findings deciphered the molecular mechanisms underlying the regulation of different HRs, from the extracellular ligand-binding pockets and transmembrane motifs to the intracellular G protein coupling interfaces. These insights are expected to facilitate selective drug discovery targeting HRs for diverse therapeutic purposes. |
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| spelling | doaj-art-41449d27401e400db59cb6878fddc55b2025-08-20T01:48:45ZengMDPI AGPharmaceuticals1424-82472025-02-0118329210.3390/ph18030292Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein BiasAnqi Chen0Chenxi Su1Zisu Zhang2Haitao Zhang3The Second Affiliated Hospital of Zhejiang University School of Medicine, Research Center for Clinical Pharmacy, Key Laboratory of Neuropharmacology and Translational Medicine of Zhejiang Province, State Key Laboratory of Advanced Drug Delivery and Release Systems, Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaThe Second Affiliated Hospital of Zhejiang University School of Medicine, Research Center for Clinical Pharmacy, Key Laboratory of Neuropharmacology and Translational Medicine of Zhejiang Province, State Key Laboratory of Advanced Drug Delivery and Release Systems, Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaThe Second Affiliated Hospital of Zhejiang University School of Medicine, Research Center for Clinical Pharmacy, Key Laboratory of Neuropharmacology and Translational Medicine of Zhejiang Province, State Key Laboratory of Advanced Drug Delivery and Release Systems, Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, ChinaThe Second Affiliated Hospital of Zhejiang University School of Medicine, Research Center for Clinical Pharmacy, Key Laboratory of Neuropharmacology and Translational Medicine of Zhejiang Province, State Key Laboratory of Advanced Drug Delivery and Release Systems, Institute of Pharmacology and Toxicology, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China<b>Background:</b> The four subtypes of G protein-coupled receptors (GPCRs) regulated by histamine play critical roles in various physiological and pathological processes, such as allergy, gastric acid secretion, cognitive and sleep disorders, and inflammation. Previous experimental structures of histamine receptors (HRs) with agonists and antagonists exhibited multiple conformations for the ligands and G protein binding. However, the structural basis for HR regulation and signaling remains elusive. <b>Methods:</b> We determined the cryo-electron microscopy (cryo-EM) structure of the H4R-histamine-Gi complex at 2.9 Å resolution, and predicted the models for all four HRs in the ligand-free apo and G protein subtype binding states using AlphaFold3 (AF3). <b>Results:</b> By comparing our H4R structure with the experimental HR structures and the computational AF3 models, we elucidated the distinct histamine binding modes and G protein interfaces, and proposed the essential roles of Y<sup>6.51</sup> and Q<sup>7.42</sup> in receptor activation and the intracellular loop 2 (ICL2) in G protein bias. <b>Conclusions:</b> Our findings deciphered the molecular mechanisms underlying the regulation of different HRs, from the extracellular ligand-binding pockets and transmembrane motifs to the intracellular G protein coupling interfaces. These insights are expected to facilitate selective drug discovery targeting HRs for diverse therapeutic purposes.https://www.mdpi.com/1424-8247/18/3/292histamine receptorsligand-binding pocketactivation mechanismG protein biascryo-EMAlphaFold3 |
| spellingShingle | Anqi Chen Chenxi Su Zisu Zhang Haitao Zhang Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein Bias Pharmaceuticals histamine receptors ligand-binding pocket activation mechanism G protein bias cryo-EM AlphaFold3 |
| title | Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein Bias |
| title_full | Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein Bias |
| title_fullStr | Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein Bias |
| title_full_unstemmed | Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein Bias |
| title_short | Cryo-EM Structures and AlphaFold3 Models of Histamine Receptors Reveal Diverse Ligand Binding and G Protein Bias |
| title_sort | cryo em structures and alphafold3 models of histamine receptors reveal diverse ligand binding and g protein bias |
| topic | histamine receptors ligand-binding pocket activation mechanism G protein bias cryo-EM AlphaFold3 |
| url | https://www.mdpi.com/1424-8247/18/3/292 |
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