Peptidyl-Resin Substrates as a Tool in the Analysis of Caspase Activity.

ABSTRACT: Caspases, proteolytic enzymes belonging to the group of cysteine proteases, play a crucial role in apoptosis. Understanding their activity and substrate specificity is extremely important. Fluorescence-based approaches, including fluorogenic substrates, are generally used to confirm cleavage preferences. Here we present a new method of substrate specificity and activity analysis based on the application of fix-charge tagged peptides located on the resin. The proteolysis of peptide bond on the resin, occurring even with low efficiency, results in the formation of N-terminal fragments of model peptide containing ionization enhancers in the form of quaternary ammonium groups, allowing for ultrasensitive and reliable analysis by LC-MS/MS. The possibility of application of the proposed s

SUBMITTER: Bachor R 

PROVIDER: S-EPMC9268085 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature