De novo design of protein minibinder agonists of TLR3
Abstract Toll-like Receptor 3 (TLR3) is a pattern recognition receptor that initiates antiviral immune responses upon binding double-stranded RNA (dsRNA). Several nucleic acid-based TLR3 agonists have been explored clinically as vaccine adjuvants in cancer and infectious disease, but present substan...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56369-w |
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| author | Chloe S. Adams Hyojin Kim Abigail E. Burtner Dong Sun Lee Craig Dobbins Cameron Criswell Brian Coventry Adri Tran-Pearson Ho Min Kim Neil P. King |
| author_facet | Chloe S. Adams Hyojin Kim Abigail E. Burtner Dong Sun Lee Craig Dobbins Cameron Criswell Brian Coventry Adri Tran-Pearson Ho Min Kim Neil P. King |
| author_sort | Chloe S. Adams |
| collection | DOAJ |
| description | Abstract Toll-like Receptor 3 (TLR3) is a pattern recognition receptor that initiates antiviral immune responses upon binding double-stranded RNA (dsRNA). Several nucleic acid-based TLR3 agonists have been explored clinically as vaccine adjuvants in cancer and infectious disease, but present substantial manufacturing and formulation challenges. Here, we use computational protein design to create novel miniproteins that bind to human TLR3 with nanomolar affinities. Cryo-EM structures of two minibinders in complex with TLR3 reveal that they bind the target as designed, although one partially unfolds due to steric competition with a nearby N-linked glycan. Multivalent forms of both minibinders induce NF-κB signaling in TLR3-expressing cell lines, demonstrating that they may have therapeutically relevant biological activity. Our work provides a foundation for the development of specific, stable, and easy-to-formulate protein-based agonists of TLRs and other pattern recognition receptors. |
| format | Article |
| id | doaj-art-04571a2c2776491db8e744f8b648572e |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-04571a2c2776491db8e744f8b648572e2025-02-02T12:32:32ZengNature PortfolioNature Communications2041-17232025-01-0116111110.1038/s41467-025-56369-wDe novo design of protein minibinder agonists of TLR3Chloe S. Adams0Hyojin Kim1Abigail E. Burtner2Dong Sun Lee3Craig Dobbins4Cameron Criswell5Brian Coventry6Adri Tran-Pearson7Ho Min Kim8Neil P. King9Institute for Protein Design, University of WashingtonCenter for Biomolecular & Cellular Structure, Institute for Basic Science (IBS)Institute for Protein Design, University of WashingtonCenter for Biomolecular & Cellular Structure, Institute for Basic Science (IBS)Institute for Protein Design, University of WashingtonInstitute for Protein Design, University of WashingtonInstitute for Protein Design, University of WashingtonInstitute for Protein Design, University of WashingtonCenter for Biomolecular & Cellular Structure, Institute for Basic Science (IBS)Institute for Protein Design, University of WashingtonAbstract Toll-like Receptor 3 (TLR3) is a pattern recognition receptor that initiates antiviral immune responses upon binding double-stranded RNA (dsRNA). Several nucleic acid-based TLR3 agonists have been explored clinically as vaccine adjuvants in cancer and infectious disease, but present substantial manufacturing and formulation challenges. Here, we use computational protein design to create novel miniproteins that bind to human TLR3 with nanomolar affinities. Cryo-EM structures of two minibinders in complex with TLR3 reveal that they bind the target as designed, although one partially unfolds due to steric competition with a nearby N-linked glycan. Multivalent forms of both minibinders induce NF-κB signaling in TLR3-expressing cell lines, demonstrating that they may have therapeutically relevant biological activity. Our work provides a foundation for the development of specific, stable, and easy-to-formulate protein-based agonists of TLRs and other pattern recognition receptors.https://doi.org/10.1038/s41467-025-56369-w |
| spellingShingle | Chloe S. Adams Hyojin Kim Abigail E. Burtner Dong Sun Lee Craig Dobbins Cameron Criswell Brian Coventry Adri Tran-Pearson Ho Min Kim Neil P. King De novo design of protein minibinder agonists of TLR3 Nature Communications |
| title | De novo design of protein minibinder agonists of TLR3 |
| title_full | De novo design of protein minibinder agonists of TLR3 |
| title_fullStr | De novo design of protein minibinder agonists of TLR3 |
| title_full_unstemmed | De novo design of protein minibinder agonists of TLR3 |
| title_short | De novo design of protein minibinder agonists of TLR3 |
| title_sort | de novo design of protein minibinder agonists of tlr3 |
| url | https://doi.org/10.1038/s41467-025-56369-w |
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