A common allosteric site and mechanism in caspases

• DOI: 10.1073/pnas.0602571103
• Publication Date: 2006-05-09T01:41:15Z
• Authors: Justin M. Scheer; Michael J. Romano...; James A. Wells
• Keywords: Inflammation; Models, Molecular; 0303 health sciences; Binding Sites; Molecular Structure; Protein Conformation; Molecular Sequence Data; Apoptosis; Crystallography, X-Ray; Caspase Inhibitors; 03 medical and health sciences; Allosteric Regulation; Caspases; Humans; Amino Acid Sequence; Disulfides; Sulfhydryl Compounds; Sequence Alignment

We present a common allosteric mechanism for control of inflammatory and apoptotic caspases. Highly specific thiol-containing inhibitors of the human inflammatory caspase-1 were identified by using disulfide trapping, a method for site-directed small-molecule discovery. These compounds became trapped by forming a disulfide bond with a cysteine residue in the cavity at the dimer interface ≈15 Å away from the active site. Mutational and structural analysis uncovered a linear circuit of functional residues that runs from one active site through the allosteric cavity and into the second active site. Kinetic analysis revealed robust positive cooperativity not seen in other endopeptidases. Recently, disulfide trapping identified a similar small-molecule site and allosteric transition in the apoptotic caspase-7 that shares only a 23% sequence identity with caspase-1. Together, these studies show a general small-molecule-binding site for functionally reversing the zymogen activation of caspases and suggest a common regulatory site for the allosteric control of inflammation and apoptosis.