Fluoxetine targets an allosteric site in the enterovirus 2C AAA+ ATPase and stabilizes a ring-shaped hexameric complex
Daniel L. Hurdiss, Priscila El Kazzi, Lisa Bauer, Nicolas Papageorgiou, François Ferrón, Tim Donselaar, Arno L. W. van Vliet, Tatiana M. Shamorkina, Joost Snijder, Bruno Canard, Étienne Decroly, Andrea Brancale, Tzviya Zeev‐Ben‐Mordehai, Friedrich Förster, Frank J. M. van Kuppeveld, Bruno Coutard
Abstract
)-fluoxetine (SFX), revealing an allosteric binding site. To study the functional consequences of SFX binding, we engineered an adenosine triphosphatase (ATPase)–competent, hexameric 2C protein. Using this system, we show that SFX, dibucaine, HBB [2-(α-hydroxybenzyl)-benzimidazole], and guanidine hydrochloride inhibit 2C ATPase activity. Moreover, cryo–electron 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.