Figure 5.

Compound 6 interacts with pro-Caspase-1 CARD to block its cleavage. (A) Pyroptosome assay showed compound 6 blocks ASC mediated caspase-1 cleavage. (B and C) DARTS assay revealed compound 6 protects the caspase-1 from proteolysis and this protection is relying on the CARD domain. (A, B, and C) Representative gels from n ≥ 3 independent experiments. (B and C) Histone 3 protein served as a loading indicator. (D) Surface docking model of compound 6–CARD complex; compound 6 is green. Enlarged image showing the details of interaction. The yellow and blue dashed lines indicate the hydrogen bonds. (C and D) human caspase-1 CARD, PDB ID: 5FNA. (E) Predicted molecular bonds and distance of residues between compound 6 and caspase-1. (F) Protein sequence alignment of the N-terminal of caspase proteins. Asterisks (*) indicates the amino acids potentially interacting with compound 6. Gray shadowed sequences are CARD.