Structural studies of the IFNλ4 receptor complex using cryoEM enabled by protein engineering
Abstract IFNλ4 has posed a conundrum in human immunology since its discovery in 2013, with its expression linked to complications with viral clearance. While genetic and cellular studies revealed the detrimental effects of IFNλ4 expression, extensive structural and functional characterization has be...
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| Format: | Article |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56119-y |
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| author | William S. Grubbe Bixia Zhang Aileen Kauffman Fabian Byléhn Kasia Padoł Hae-Gwang Jung Seung Bum Park Jessica M. Priest Engin Özkan Juan J. de Pablo T. Jake Liang Minglei Zhao Juan L. Mendoza |
| author_facet | William S. Grubbe Bixia Zhang Aileen Kauffman Fabian Byléhn Kasia Padoł Hae-Gwang Jung Seung Bum Park Jessica M. Priest Engin Özkan Juan J. de Pablo T. Jake Liang Minglei Zhao Juan L. Mendoza |
| author_sort | William S. Grubbe |
| collection | DOAJ |
| description | Abstract IFNλ4 has posed a conundrum in human immunology since its discovery in 2013, with its expression linked to complications with viral clearance. While genetic and cellular studies revealed the detrimental effects of IFNλ4 expression, extensive structural and functional characterization has been limited by the inability to express and purify the protein, complicating explanations of its paradoxical behavior. In this work, we report a method for robust production of IFNλ4. We then use yeast surface display to affinity-mature IL10Rβ and solve the 72 kilodalton structures of IFNλ4 (3.26 Å) and IFNλ3 (3.00 Å) in complex with their receptors IFNλR1 and IL10Rβ using cryogenic electron microscopy. Comparison of the structures highlights differences in receptor engagement and reveals a distinct 12-degree rotation in overall receptor geometry, providing a potential mechanistic explanation for differences in cell signaling, downstream gene induction, and antiviral activities. Further, we perform a structural analysis using molecular modeling and simulation to identify a unique region of IFNλ4 that, when replaced, enables secretion of the protein from cells. These findings provide a structural and functional understanding of the IFNλ4 protein and enable future comprehensive studies towards correcting IFNλ4 dysfunction in large populations of affected patients. |
| format | Article |
| id | doaj-art-ad38ac86c40e4053a4000170820f700a |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-ad38ac86c40e4053a4000170820f700a2025-01-19T12:31:03ZengNature PortfolioNature Communications2041-17232025-01-0116111210.1038/s41467-025-56119-yStructural studies of the IFNλ4 receptor complex using cryoEM enabled by protein engineeringWilliam S. Grubbe0Bixia Zhang1Aileen Kauffman2Fabian Byléhn3Kasia Padoł4Hae-Gwang Jung5Seung Bum Park6Jessica M. Priest7Engin Özkan8Juan J. de Pablo9T. Jake Liang10Minglei Zhao11Juan L. Mendoza12Pritzker School of Molecular Engineering, University of ChicagoDepartment of Biochemistry and Molecular Biology, University of ChicagoPritzker School of Molecular Engineering, University of ChicagoPritzker School of Molecular Engineering, University of ChicagoPritzker School of Molecular Engineering, University of ChicagoLiver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of HealthLiver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of HealthDepartment of Biochemistry and Molecular Biology, University of ChicagoDepartment of Biochemistry and Molecular Biology, University of ChicagoPritzker School of Molecular Engineering, University of ChicagoLiver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of HealthDepartment of Biochemistry and Molecular Biology, University of ChicagoPritzker School of Molecular Engineering, University of ChicagoAbstract IFNλ4 has posed a conundrum in human immunology since its discovery in 2013, with its expression linked to complications with viral clearance. While genetic and cellular studies revealed the detrimental effects of IFNλ4 expression, extensive structural and functional characterization has been limited by the inability to express and purify the protein, complicating explanations of its paradoxical behavior. In this work, we report a method for robust production of IFNλ4. We then use yeast surface display to affinity-mature IL10Rβ and solve the 72 kilodalton structures of IFNλ4 (3.26 Å) and IFNλ3 (3.00 Å) in complex with their receptors IFNλR1 and IL10Rβ using cryogenic electron microscopy. Comparison of the structures highlights differences in receptor engagement and reveals a distinct 12-degree rotation in overall receptor geometry, providing a potential mechanistic explanation for differences in cell signaling, downstream gene induction, and antiviral activities. Further, we perform a structural analysis using molecular modeling and simulation to identify a unique region of IFNλ4 that, when replaced, enables secretion of the protein from cells. These findings provide a structural and functional understanding of the IFNλ4 protein and enable future comprehensive studies towards correcting IFNλ4 dysfunction in large populations of affected patients.https://doi.org/10.1038/s41467-025-56119-y |
| spellingShingle | William S. Grubbe Bixia Zhang Aileen Kauffman Fabian Byléhn Kasia Padoł Hae-Gwang Jung Seung Bum Park Jessica M. Priest Engin Özkan Juan J. de Pablo T. Jake Liang Minglei Zhao Juan L. Mendoza Structural studies of the IFNλ4 receptor complex using cryoEM enabled by protein engineering Nature Communications |
| title | Structural studies of the IFNλ4 receptor complex using cryoEM enabled by protein engineering |
| title_full | Structural studies of the IFNλ4 receptor complex using cryoEM enabled by protein engineering |
| title_fullStr | Structural studies of the IFNλ4 receptor complex using cryoEM enabled by protein engineering |
| title_full_unstemmed | Structural studies of the IFNλ4 receptor complex using cryoEM enabled by protein engineering |
| title_short | Structural studies of the IFNλ4 receptor complex using cryoEM enabled by protein engineering |
| title_sort | structural studies of the ifnλ4 receptor complex using cryoem enabled by protein engineering |
| url | https://doi.org/10.1038/s41467-025-56119-y |
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