Differentiating the Substrate Profiles of Inflammatory Caspases Using Extended Forster Resonance Energy Transfer-Based Peptide Substrates.

ABSTRACT: Human inflammatory caspases (caspase-1, -4, and -5) are key players in the innate immune response. These enzymes have been shown to cleave proinflammatory substrates, implicating them in many inflammatory disease states. Their activity is frequently assessed using in vitro fluorogenic assays, with all three human inflammatory caspases preferring the same WEHD tetrapeptide. The study examines the specificity of these enzymes C-terminal to the cleaved aspartate residue with Förster resonance energy transfer peptide-based assays using 7-methoxycoumaryl alanine [A(MCA)] as the donor and lysine-conjugated dabsyl [K(Dab)] as the quencher. The P₄-P₁ peptide sequences A(MCA)EHD, A(MCA)VAD, and A(MCA)QPD are varied on the C-terminal (prime) side of the peptide. Historically, c

SUBMITTER: Boyd TC 

PROVIDER: S-EPMC12695177 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature