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 water to react with the acyl-enzyme intermediate.There have been several examples of protease-mediated transpeptidation, but they are generally inefficient, and Brake Assembly 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 to the C-termini of Books 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.