Identification of Nucleophilic Probes for Protease-Mediated Transpeptidation

Identification of Nucleophilic Probes for Protease-Mediated Transpeptidation

Blog Article

Proteases have evolved to mediate the hydrolysis of peptide bonds but may perform transpeptidation in the presence of a proper nucleophilic molecule that can effectively compete with Cleansing Sponge water to react with the acyl-enzyme intermediate.There have been several examples of protease-mediated transpeptidation, but they are generally inefficient, and little effort has been made to systematically control the transpeptidation activity of other proteases with good nucleophiles.Here, we developed an on-bead screening approach to find a probe that functions efficiently as a nucleophile in the protease-mediated transpeptidation reaction, and we identified good probes for a model protease DegP.These probes were covalently linked Cover to the C-termini of the cleaved peptides in a mild condition and made the selective enrichment of ligated peptides possible.We suggest that good nucleophilic probes can be found for many other proteases that act via acyl-enzyme intermediates, and these probes will help characterize their substrates.