A coupled protein and probe engineering approach for selective inhibition and activity-based probe labeling of the caspases.

ABSTRACT: Caspases are cysteine proteases that play essential roles in apoptosis and inflammation. Unfortunately, their highly conserved active sites and overlapping substrate specificities make it difficult to use inhibitors or activity-based probes to study the function, activation, localization, and regulation of individual members of this family. Here we describe a strategy to engineer a caspase to contain a latent nucleophile that can be targeted by a probe containing a suitably placed electrophile, thereby allowing specific, irreversible inhibition and labeling of only the engineered protease. To accomplish this, we have identified a non-conserved residue on the small subunit of all caspases that is near the substrate-binding pocket and that can be mutated to a non-catalytic cysteine residue.

SUBMITTER: Xiao J 

PROVIDER: S-EPMC3722599 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature