Improved Asparaginyl-Ligase-Catalyzed Transpeptidation via Selective Nucleophile Quenching.

Angewandte Chemie
Fabian B H RehmDavid J Craik

Abstract

The use of enzymes for the site-specific modification of proteins/peptides has become a highly accessible, widespread approach to study protein/peptide functions or to generate therapeutic conjugates. Asparaginyl endopeptidases (AEPs) that preferentially catalyze transpeptidation reactions (AEP ligases) have emerged as enticing alternatives to established approaches, such as bacterial sortases, due to their catalytic efficiency and short tripeptide recognition motifs. However, under standard conditions, a substantial excess of the nucleophile to be conjugated is needed to reach desirable yields. Herein we report a versatile approach to shift the AEP-catalyzed transpeptidation equilibrium toward product formation via selectively quenching the nucleophilicity of the competing leaving-group peptide. Our metal-complexation-based strategy enables efficient peptide/protein labeling at the N- or C-terminus with near-equimolar concentrations of nucleophile label. Furthermore, we show that this approach can enhance protein-protein ligation and facilitate the formation of transpeptidation products that are otherwise unattainable.

References

Sep 25, 2007·Nature Chemical Biology·Maximilian W PoppHidde L Ploegh
Aug 29, 2009·Angewandte Chemie·Ellen M Sletten, Carolyn R Bertozzi
Jul 21, 2014·Nature Chemical Biology·Giang K T NguyenJames P Tam
Feb 24, 2015·Chemical Reviews·Omar Boutureira, Gonçalo J L Bernardes
Jul 15, 2015·Chemical Communications : Chem Comm·R David RowJohn M Antos
Dec 19, 2015·Nature Communications·Karen S HarrisMarilyn A Anderson
Feb 16, 2017·Journal of the American Chemical Society·Renliang YangBin Wu
Sep 28, 2018·Chemical Society Reviews·Yi ZhangMark D Distefano
Apr 5, 2019·Proceedings of the National Academy of Sciences of the United States of America·Fabian B H RehmDavid J Craik
Oct 2, 2019·Journal of the American Chemical Society·Fabian B H RehmHidde L Ploegh
Sep 3, 2020·Chemical Science·T M Simon TangLouis Y P Luk

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Citations

Mar 6, 2021·Biochemical Society Transactions·Samuel G NonisJoshua S Mylne
Mar 10, 2021·Chembiochem : a European Journal of Chemical Biology·Fabian B H RehmDavid J Craik
May 27, 2021·Organic & Biomolecular Chemistry·T M Simon Tang, Louis Y P Luk
Jun 16, 2021·ACS Chemical Biology·Thibault J HarmandHidde L Ploegh
Nov 12, 2021·Journal of the American Chemical Society·Fabian B H RehmThomas Durek

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