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Identification of Nucleophilic Probes for Protease-Mediated Transpeptidation.

ABSTRACT: 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 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 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.

SUBMITTER: Eom GE

PROVIDER: S-EPMC6225416 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature

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Identification of Nucleophilic Probes for Protease-Mediated Transpeptidation.

Eom Ga-Eul GE Kim Seokhee S

Molecules (Basel, Switzerland) 20180822 9

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 little effort has been made to systematically control the transpeptidation activity of other proteases with good nucleophiles. Here, we developed an on-bea ...[more]

PMID: 30131476

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