TY - JOUR T1 - Cryo-EM structure of the NLRP3 decamer bound to the cytokine release inhibitory drug CRID3 JF - bioRxiv DO - 10.1101/2021.07.22.453353 SP - 2021.07.22.453353 AU - Inga V. Hochheiser AU - Michael Pilsl AU - Gregor Hagelueken AU - Jonas Moecking AU - Michael Marleaux AU - Rebecca Brinkschulte AU - Eicke Latz AU - Christoph Engel AU - Matthias Geyer Y1 - 2021/01/01 UR - http://biorxiv.org/content/early/2021/07/22/2021.07.22.453353.abstract N2 - NLRP3 is an intracellular sensor protein whose activation by a broad spectrum of exogenous and endogenous stimuli leads to inflammasome formation and pyroptosis. The mechanisms leading to NLRP3 activation and the way how antagonistic small molecules function remain poorly understood. Here we report the cryo-electron microscopy structures of full-length NLRP3 in its native form and complexed with the inhibitor CRID3 (also named MCC950). Inactive, ADP-bound NLRP3 is a decamer composed of homodimers of intertwined LRR domains that assemble back-to-back as pentamers with the NACHT domain located at the apical axis of this spherical structure. Molecular contacts between the concave sites of two opposing LRRs are mediated by an acidic loop extending from an LRR transition segment. Binding of CRID3 significantly stabilizes the NACHT and LRR domains relative to each other, allowing structural resolution of 3.9-4.2 Å. CRID3 binds into a cleft, connecting four subdomains of the NACHT with the transition LRR. Its central sulfonylurea group interacts with the Walker A motif of the NLRP3 nucleotide-binding domain and is sandwiched between two arginines from opposing sites, explaining the specificity of NLRP3 for this chemical entity. With the determination of the binding site of this lead therapeutic, specific targeting of NLRP3 for the treatment of autoinflammatory and autoimmune diseases and rational drug optimization are within reach.Competing Interest StatementM.G. and E.L. are co-founders and consultants of IFM Therapeutics. The other authors declare no competing interests. ER -