Leveraging Sortase A Electrostatics for Powerful Transpeptidation Reactions
Sortase-mediated transpeptidation is a powerful biochemical reaction to perform protein engineering. In this work, we leverage the unique electrostatic profile of sortase A pentamutant (SrtA-5M) to improve SrtA-5M-mediated transpeptidations by incorporating short, charged peptidic modules into the substrates. Importantly, the reaction proceeds with a minimal excess of nucleophile and is fast and highly efficient in the low micromolar substrate concentration range. Electrostatic assistance eliminates the need for additives or complex substrate engineering strategies, thereby giving it a broad scope. Our findings also provide fundamental insights into the influence of substrate charge on SrtA-5M activity, paving the way for further optimization of sortase A-catalyzed transpeptidation reactions.
Authors
- Benoît Snella
- Jérôme Vicogne (ORCID: https://orcid.org/0000-0001-8360-7497)
- Vangelis Agouridas (ORCID: https://orcid.org/0000-0003-0911-1527)
- Oleg Melnyk (ORCID: https://orcid.org/0000-0002-3863-5613)
- Chen Wang (ORCID: https://orcid.org/0000-0003-1963-4954)
- Rémi Desmet (ORCID: https://orcid.org/0000-0002-4150-7399)
Institutions
- Centre National de la Recherche Scientifique (FR)
- École Nationale Supérieure de Chimie de Lille (FR)
- Inserm (FR)
- Université de Lille (FR)
- Institut Pasteur de Lille (FR)
- Centre Hospitalier Universitaire de Lille (FR)
- Center for Infection and Immunity of Lille (FR)
- École Centrale de Lille (FR)
- University of the Arts London (GB)
Publication Details
- Journal
- Angewandte Chemie International Edition
- Published
- 2025-05-09
- DOI
- https://doi.org/10.1002/anie.202507236
- Citations
- 3
- Primary Topic
- Biochemical and Structural Characterization
- Type
- article
- Field-Weighted Citation Impact
- 2.42
Funders
- Université de Lille
- Agence Nationale de la Recherche
- Institut National de la Santé et de la Recherche Médicale
- Austrian Science Fund
- Centre National de la Recherche Scientifique