Structural Analysis of Inhibitor Binding to Enterovirus-D68 3C Protease
Enterovirus-D68 (EV68) continues to present as a global health issue causing respiratory illness and outbreaks associated with long-lasting neurological disease, with no antivirals or specific treatment options. The development of antiviral therapeutics, such as small-molecule inhibitors that target...
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MDPI AG
2025-01-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/17/1/75 |
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| author | Vincent N. Azzolino Ala M. Shaqra Akbar Ali Nese Kurt Yilmaz Celia A. Schiffer |
| author_facet | Vincent N. Azzolino Ala M. Shaqra Akbar Ali Nese Kurt Yilmaz Celia A. Schiffer |
| author_sort | Vincent N. Azzolino |
| collection | DOAJ |
| description | Enterovirus-D68 (EV68) continues to present as a global health issue causing respiratory illness and outbreaks associated with long-lasting neurological disease, with no antivirals or specific treatment options. The development of antiviral therapeutics, such as small-molecule inhibitors that target conserved proteins like the enteroviral 3C protease, remains to be achieved. While various 3C inhibitors have been investigated, their design does not consider the potential emergence of drug resistance mutations. For other antivirals where resistance has been a challenge, we have demonstrated that the likelihood of resistance can be minimized by designing inhibitors that leverage the evolutionary constraints of the target. Here, we characterize a series of 3C inhibitors against EV68-3C protease through enzyme inhibition, protein crystallography, and structural analysis. We have determined and analyzed three high-resolution inhibitor-bound crystal structures of EV68-3C protease, which revealed possible sites of resistance mutations, a key structural water molecule conserved during ligand binding, and the conformational flexibility of the catalytic histidine H40. This structural analysis combined with enzymatic assays provides insights for the rational design of inhibitors that are robust against resistance toward developing antiviral treatments for EV68 infections. |
| format | Article |
| id | doaj-art-b5bc87f35eb644c6ab7a40941777d3d6 |
| institution | Kabale University |
| issn | 1999-4915 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Viruses |
| spelling | doaj-art-b5bc87f35eb644c6ab7a40941777d3d62025-01-24T13:52:29ZengMDPI AGViruses1999-49152025-01-011717510.3390/v17010075Structural Analysis of Inhibitor Binding to Enterovirus-D68 3C ProteaseVincent N. Azzolino0Ala M. Shaqra1Akbar Ali2Nese Kurt Yilmaz3Celia A. Schiffer4Department of Biochemistry and Molecular Biotechnology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USADepartment of Biochemistry and Molecular Biotechnology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USADepartment of Biochemistry and Molecular Biotechnology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USADepartment of Biochemistry and Molecular Biotechnology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USADepartment of Biochemistry and Molecular Biotechnology, University of Massachusetts Chan Medical School, Worcester, MA 01605, USAEnterovirus-D68 (EV68) continues to present as a global health issue causing respiratory illness and outbreaks associated with long-lasting neurological disease, with no antivirals or specific treatment options. The development of antiviral therapeutics, such as small-molecule inhibitors that target conserved proteins like the enteroviral 3C protease, remains to be achieved. While various 3C inhibitors have been investigated, their design does not consider the potential emergence of drug resistance mutations. For other antivirals where resistance has been a challenge, we have demonstrated that the likelihood of resistance can be minimized by designing inhibitors that leverage the evolutionary constraints of the target. Here, we characterize a series of 3C inhibitors against EV68-3C protease through enzyme inhibition, protein crystallography, and structural analysis. We have determined and analyzed three high-resolution inhibitor-bound crystal structures of EV68-3C protease, which revealed possible sites of resistance mutations, a key structural water molecule conserved during ligand binding, and the conformational flexibility of the catalytic histidine H40. This structural analysis combined with enzymatic assays provides insights for the rational design of inhibitors that are robust against resistance toward developing antiviral treatments for EV68 infections.https://www.mdpi.com/1999-4915/17/1/75enterovirusEV68proteasedrug resistanceprotein structureinhibition assays |
| spellingShingle | Vincent N. Azzolino Ala M. Shaqra Akbar Ali Nese Kurt Yilmaz Celia A. Schiffer Structural Analysis of Inhibitor Binding to Enterovirus-D68 3C Protease Viruses enterovirus EV68 protease drug resistance protein structure inhibition assays |
| title | Structural Analysis of Inhibitor Binding to Enterovirus-D68 3C Protease |
| title_full | Structural Analysis of Inhibitor Binding to Enterovirus-D68 3C Protease |
| title_fullStr | Structural Analysis of Inhibitor Binding to Enterovirus-D68 3C Protease |
| title_full_unstemmed | Structural Analysis of Inhibitor Binding to Enterovirus-D68 3C Protease |
| title_short | Structural Analysis of Inhibitor Binding to Enterovirus-D68 3C Protease |
| title_sort | structural analysis of inhibitor binding to enterovirus d68 3c protease |
| topic | enterovirus EV68 protease drug resistance protein structure inhibition assays |
| url | https://www.mdpi.com/1999-4915/17/1/75 |
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