Design of a recombinant asparaginyl ligase for site-specific modification using efficient recognition and nucleophile motifs.

ABSTRACT: Asparaginyl ligases have been extensively utilized as valuable tools for site-specific bioconjugation or surface-modification. However, the application is hindered by the laborious and poorly reproducible preparation processes, unstable activity and ambiguous substrate requirements. To address these limitations, this study employed a structure-based rational approach to obtain a high-yield and high-activity protein ligase called OaAEP1-C247A-aa55-351. It was observed that OaAEP1-C247A-aa55-351 exhibits appreciable catalytic activities across a wide pH range, and the addition of the Fe³⁺ metal ion effectively enhances the catalytic power. Importantly, this study provides insight into the recognition and nucleophile peptide profiles of OaAEP1-C247A-aa55-351. The ligase demonstrate

SUBMITTER: Tang J 

PROVIDER: S-EPMC11026461 | biostudies-literature | 2024 Apr

REPOSITORIES: biostudies-literature