Molecular mechanism underlying non-discriminatory recognition of relaxin-3 by RXFP3 and RXFP4

Abstract The human relaxin family peptide receptors RXFP3 and RXFP4 play important physiological roles through interactions with endogenous hormones, relaxin-3 and insulin-like peptide 5 (INSL5). They are implicated in certain neurological and metabolic disorders. While INSL5 only activates RXFP4, r...

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Main Authors: Yan Chen, Qingtong Zhou, Shiyu Yan, Jiahui Yan, Dehua Yang, Jian Chen, Ming-Wei Wang
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Communications Biology
Online Access:https://doi.org/10.1038/s42003-025-08220-7
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author Yan Chen
Qingtong Zhou
Shiyu Yan
Jiahui Yan
Dehua Yang
Jian Chen
Ming-Wei Wang
author_facet Yan Chen
Qingtong Zhou
Shiyu Yan
Jiahui Yan
Dehua Yang
Jian Chen
Ming-Wei Wang
author_sort Yan Chen
collection DOAJ
description Abstract The human relaxin family peptide receptors RXFP3 and RXFP4 play important physiological roles through interactions with endogenous hormones, relaxin-3 and insulin-like peptide 5 (INSL5). They are implicated in certain neurological and metabolic disorders. While INSL5 only activates RXFP4, relaxin-3 is recognized by both receptors. Here, we report the cryo-electron microscopy structures of RXFP3–Gi complexes bound by relaxin-3 or a small-molecule dual agonist (compound 4), and relaxin-3 in complex with RXFP4–Gi, with global resolutions of 2.91 Å, 2.95 Å, and 3.10 Å, respectively. It is found that relaxin-3 adopts a conserved binding conformation within the transmembrane domain (TMD) bundle of RXFP3 and RXFP4, where the C-terminal tip residues of its B chain, R26 and W27, make extensive contacts with conserved receptor residues, thereby activating RXFP3 and RXFP4. Compound 4 mimics these key interactions by binding to both receptors. In contrast, the C-terminal residues composition and TMD-binding angle of INSL5 in RXFP4 differ significantly from that of relaxin-3, ensuring its selectivity for RXFP4. These findings deepen our understanding about the structural basis of ligand recognition and selectivity in this G protein-coupled receptor subfamily.
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issn 2399-3642
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spelling doaj-art-5437befc0e5c4fa2a68fd42b1d63b9572025-08-20T03:48:15ZengNature PortfolioCommunications Biology2399-36422025-05-018111310.1038/s42003-025-08220-7Molecular mechanism underlying non-discriminatory recognition of relaxin-3 by RXFP3 and RXFP4Yan Chen0Qingtong Zhou1Shiyu Yan2Jiahui Yan3Dehua Yang4Jian Chen5Ming-Wei Wang6Research Center for Medicinal Structural Biology, National Research Center for Translational Medicine at Shanghai, State Key Laboratory of Medical Genomics, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineResearch Center for Medicinal Structural Biology, National Research Center for Translational Medicine at Shanghai, State Key Laboratory of Medical Genomics, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineThe CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesThe CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesThe CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesSchool of Pharmacy, Fudan UniversityResearch Center for Medicinal Structural Biology, National Research Center for Translational Medicine at Shanghai, State Key Laboratory of Medical Genomics, Ruijin Hospital, Shanghai Jiao Tong University School of MedicineAbstract The human relaxin family peptide receptors RXFP3 and RXFP4 play important physiological roles through interactions with endogenous hormones, relaxin-3 and insulin-like peptide 5 (INSL5). They are implicated in certain neurological and metabolic disorders. While INSL5 only activates RXFP4, relaxin-3 is recognized by both receptors. Here, we report the cryo-electron microscopy structures of RXFP3–Gi complexes bound by relaxin-3 or a small-molecule dual agonist (compound 4), and relaxin-3 in complex with RXFP4–Gi, with global resolutions of 2.91 Å, 2.95 Å, and 3.10 Å, respectively. It is found that relaxin-3 adopts a conserved binding conformation within the transmembrane domain (TMD) bundle of RXFP3 and RXFP4, where the C-terminal tip residues of its B chain, R26 and W27, make extensive contacts with conserved receptor residues, thereby activating RXFP3 and RXFP4. Compound 4 mimics these key interactions by binding to both receptors. In contrast, the C-terminal residues composition and TMD-binding angle of INSL5 in RXFP4 differ significantly from that of relaxin-3, ensuring its selectivity for RXFP4. These findings deepen our understanding about the structural basis of ligand recognition and selectivity in this G protein-coupled receptor subfamily.https://doi.org/10.1038/s42003-025-08220-7
spellingShingle Yan Chen
Qingtong Zhou
Shiyu Yan
Jiahui Yan
Dehua Yang
Jian Chen
Ming-Wei Wang
Molecular mechanism underlying non-discriminatory recognition of relaxin-3 by RXFP3 and RXFP4
Communications Biology
title Molecular mechanism underlying non-discriminatory recognition of relaxin-3 by RXFP3 and RXFP4
title_full Molecular mechanism underlying non-discriminatory recognition of relaxin-3 by RXFP3 and RXFP4
title_fullStr Molecular mechanism underlying non-discriminatory recognition of relaxin-3 by RXFP3 and RXFP4
title_full_unstemmed Molecular mechanism underlying non-discriminatory recognition of relaxin-3 by RXFP3 and RXFP4
title_short Molecular mechanism underlying non-discriminatory recognition of relaxin-3 by RXFP3 and RXFP4
title_sort molecular mechanism underlying non discriminatory recognition of relaxin 3 by rxfp3 and rxfp4
url https://doi.org/10.1038/s42003-025-08220-7
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