Proximity-Based Sortase-Mediated Ligation

Overview

Journal Angew Chem Int Ed Engl

Specialty Chemistry

Date 2017 Apr 5

PMID 28374553

Citations 29

Authors

Hejia Henry Wang

Burcin Altun

Kido Nwe

Andrew Tsourkas

Affiliations

Soon will be listed here.

Abstract

Protein bioconjugation has been a crucial tool for studying biological processes and developing therapeutics. Sortase A (SrtA), a bacterial transpeptidase, has become widely used for its ability to site-specifically label proteins with diverse functional moieties, but a significant limitation is its poor reaction kinetics. In this work, we address this by developing proximity-based sortase-mediated ligation (PBSL), which improves the ligation efficiency to over 95 % by linking the target protein to SrtA using the SpyTag-SpyCatcher peptide-protein pair. By expressing the target protein with SpyTag C-terminal to the SrtA recognition motif, it can be covalently captured by an immobilized SpyCatcher-SrtA fusion protein during purification. Following the ligation reaction, SpyTag is cleaved off, rendering PBSL traceless, and only the labeled protein is released, simplifying target protein purification and labeling to a single step.

Citing Articles

Development of a Versatile Cancer Vaccine Format Targeting Antigen-Presenting Cells Using Proximity-Based Sortase A-Mediated Ligation of T-Cell Epitopes.

Wang A, Brink H, Bouma R, Affandi A, Nijen Twilhaar M, Heijnen D Bioconjug Chem. 2024; 35(11):1805-1814.

PMID: 39511711 PMC: 11583207. DOI: 10.1021/acs.bioconjchem.4c00403.

Sortase mediated protein ubiquitination with defined chain length and topology.

Raniszewski N, Beyer J, Noel M, Burslem G RSC Chem Biol. 2024; 5(4):321-327.

PMID: 38576722 PMC: 10989510. DOI: 10.1039/d3cb00229b.

Intracellular Protein Delivery: Approaches, Challenges, and Clinical Applications.

Chan A, Tsourkas A BME Front. 2024; 5:0035.

PMID: 38282957 PMC: 10809898. DOI: 10.34133/bmef.0035.

Nature-inspired protein ligation and its applications.

Pihl R, Zheng Q, David Y Nat Rev Chem. 2023; 7(4):234-255.

PMID: 37117416 PMC: 10659114. DOI: 10.1038/s41570-023-00468-z.

Lipid Nanoparticle Delivery of Small Proteins for Potent RAS Inhibition.

Haley R, Chan A, Billingsley M, Gong N, Padilla M, Kim E ACS Appl Mater Interfaces. 2023; 15(18):21877-21892.

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