Distinct structural mechanisms of LGR4 modulation by Norrin and RSPOs in Wnt/β-catenin signaling

Abstract The Wnt/β-catenin pathway requires precise regulation for proper development and tissue homeostasis, yet the structural mechanisms enabling its fine-tuned control remain incompletely understood. Here, we reveal how LGR4 achieves differential signaling outcomes through distinct recognition o...

Full description

Saved in:
Bibliographic Details
Main Authors: Huarui Qiao, Fangzheng Hu, Yiang Wang, Lu Wang, Siyu Zhou, Shaojue Guo, Yiwen Xu, Jianfeng Xu, Qianqian Cui, Qilun Yang, H. Eric Xu, Jianwei Zhu, Yong Geng
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-61545-z
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849764332163825664
author Huarui Qiao
Fangzheng Hu
Yiang Wang
Lu Wang
Siyu Zhou
Shaojue Guo
Yiwen Xu
Jianfeng Xu
Qianqian Cui
Qilun Yang
H. Eric Xu
Jianwei Zhu
Yong Geng
author_facet Huarui Qiao
Fangzheng Hu
Yiang Wang
Lu Wang
Siyu Zhou
Shaojue Guo
Yiwen Xu
Jianfeng Xu
Qianqian Cui
Qilun Yang
H. Eric Xu
Jianwei Zhu
Yong Geng
author_sort Huarui Qiao
collection DOAJ
description Abstract The Wnt/β-catenin pathway requires precise regulation for proper development and tissue homeostasis, yet the structural mechanisms enabling its fine-tuned control remain incompletely understood. Here, we reveal how LGR4 achieves differential signaling outcomes through distinct recognition of two key modulators: Norrin and R-spondins (RSPOs). Using cryo-electron microscopy, we determined the structure of full-length LGR4 bound to Norrin in a 2:2 stoichiometry, revealing a molecular bridging mechanism where Norrin dimer connect two LGR4 protomers in a spatial arrangement fundamentally distinct from the LGR4-RSPO2-ZNRF3 complex. Notably, Norrin binding to LGR4 sterically hinders simultaneous interaction with the Frizzled4 receptor, establishing a regulatory checkpoint in Wnt signaling. The partially overlapping binding sites for Norrin and RSPOs on LGR4 enable mutually exclusive interactions that drive distinct signaling outcomes. Disease-linked mutations map to distinct functional regions: those disrupting LGR4 interaction are associated with familial exudative vitreoretinopathy (FEVR), while others impairing Frizzled4 binding are linked to Norrie disease. Furthermore, we developed a high-affinity nanobody that blocks both Norrin and RSPO binding to LGR4, providing a potential tool for therapeutic intervention. These findings elucidate the structural basis of LGR4’s dual signaling roles and lay the groundwork for therapeutic strategies targeting Wnt-related diseases.
format Article
id doaj-art-1c2fee0b07cc47ec80a6e1040744fbda
institution DOAJ
issn 2041-1723
language English
publishDate 2025-07-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-1c2fee0b07cc47ec80a6e1040744fbda2025-08-20T03:05:10ZengNature PortfolioNature Communications2041-17232025-07-0116111110.1038/s41467-025-61545-zDistinct structural mechanisms of LGR4 modulation by Norrin and RSPOs in Wnt/β-catenin signalingHuarui Qiao0Fangzheng Hu1Yiang Wang2Lu Wang3Siyu Zhou4Shaojue Guo5Yiwen Xu6Jianfeng Xu7Qianqian Cui8Qilun Yang9H. Eric Xu10Jianwei Zhu11Yong Geng12Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityState Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesState Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesState Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesState Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesState Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesDepartment of Biopharmaceutics, College of Food Science and Technology, Shanghai Ocean UniversityState Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesShanghai Kailuo Biotechnology Co., Ltd.State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of SciencesEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityAbstract The Wnt/β-catenin pathway requires precise regulation for proper development and tissue homeostasis, yet the structural mechanisms enabling its fine-tuned control remain incompletely understood. Here, we reveal how LGR4 achieves differential signaling outcomes through distinct recognition of two key modulators: Norrin and R-spondins (RSPOs). Using cryo-electron microscopy, we determined the structure of full-length LGR4 bound to Norrin in a 2:2 stoichiometry, revealing a molecular bridging mechanism where Norrin dimer connect two LGR4 protomers in a spatial arrangement fundamentally distinct from the LGR4-RSPO2-ZNRF3 complex. Notably, Norrin binding to LGR4 sterically hinders simultaneous interaction with the Frizzled4 receptor, establishing a regulatory checkpoint in Wnt signaling. The partially overlapping binding sites for Norrin and RSPOs on LGR4 enable mutually exclusive interactions that drive distinct signaling outcomes. Disease-linked mutations map to distinct functional regions: those disrupting LGR4 interaction are associated with familial exudative vitreoretinopathy (FEVR), while others impairing Frizzled4 binding are linked to Norrie disease. Furthermore, we developed a high-affinity nanobody that blocks both Norrin and RSPO binding to LGR4, providing a potential tool for therapeutic intervention. These findings elucidate the structural basis of LGR4’s dual signaling roles and lay the groundwork for therapeutic strategies targeting Wnt-related diseases.https://doi.org/10.1038/s41467-025-61545-z
spellingShingle Huarui Qiao
Fangzheng Hu
Yiang Wang
Lu Wang
Siyu Zhou
Shaojue Guo
Yiwen Xu
Jianfeng Xu
Qianqian Cui
Qilun Yang
H. Eric Xu
Jianwei Zhu
Yong Geng
Distinct structural mechanisms of LGR4 modulation by Norrin and RSPOs in Wnt/β-catenin signaling
Nature Communications
title Distinct structural mechanisms of LGR4 modulation by Norrin and RSPOs in Wnt/β-catenin signaling
title_full Distinct structural mechanisms of LGR4 modulation by Norrin and RSPOs in Wnt/β-catenin signaling
title_fullStr Distinct structural mechanisms of LGR4 modulation by Norrin and RSPOs in Wnt/β-catenin signaling
title_full_unstemmed Distinct structural mechanisms of LGR4 modulation by Norrin and RSPOs in Wnt/β-catenin signaling
title_short Distinct structural mechanisms of LGR4 modulation by Norrin and RSPOs in Wnt/β-catenin signaling
title_sort distinct structural mechanisms of lgr4 modulation by norrin and rspos in wnt β catenin signaling
url https://doi.org/10.1038/s41467-025-61545-z
work_keys_str_mv AT huaruiqiao distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT fangzhenghu distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT yiangwang distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT luwang distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT siyuzhou distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT shaojueguo distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT yiwenxu distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT jianfengxu distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT qianqiancui distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT qilunyang distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT hericxu distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT jianweizhu distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling
AT yonggeng distinctstructuralmechanismsoflgr4modulationbynorrinandrsposinwntbcateninsignaling