Download MendeleyFluoxetine targets an allosteric site in the enterovirus 2C AAA+ ATPase and stabilizes a ring-shaped hexameric complex.
Hurdiss, D.L., El Kazzi, P., Bauer, L., Papageorgiou, N., Ferron, F.P., Donselaar, T., van Vliet, A.L.W., Shamorkina, T.M., Snijder, J., Canard, B., Decroly, E., Brancale, A., Zeev-Ben-Mordehai, T., Forster, F., van Kuppeveld, F.J.M., Coutard, B.(2022) Sci Adv 8: eabj7615-eabj7615
- PubMed: 34985963
- DOI: https://doi.org/10.1126/sciadv.abj7615
- Primary Citation of Related Structures:
6S3A, 6T3W - PubMed Abstract:
Enteroviruses are globally prevalent human pathogens responsible for many diseases. The nonstructural protein 2C is a AAA+ helicase and plays a key role in enterovirus replication. Drug repurposing screens identified 2C-targeting compounds such as fluoxetine and dibucaine, but how they inhibit 2C is unknown. Here, we present a crystal structure of the soluble and monomeric fragment of coxsackievirus B3 2C protein in complex with ( S )-fluoxetine (SFX), revealing an allosteric binding site. To study the functional consequences of SFX binding, we engineered an adenosine triphosphatase (ATPase)¨Ccompetent, hexameric 2C protein. Using this system, we show that SFX, dibucaine, HBB [2-(¦Á-hydroxybenzyl)-benzimidazole], and guanidine hydrochloride inhibit 2C ATPase activity. Moreover, cryo¨Celectron microscopy analysis demonstrated that SFX and dibucaine lock 2C in a defined hexameric state, rationalizing their mode of inhibition. Collectively, these results provide important insights into 2C inhibition and a robust engineering strategy for structural, functional, and drug-screening analysis of 2C proteins.
Organizational Affiliation:
Virology Section, Infectious Diseases and Immunology Division, Department of Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht University, 3584CL Utrecht, Netherlands.
