Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

Use of an asparaginyl endopeptidase for chemo-enzymatic peptide and protein labeling

Tang, T. M. Simon, Cardella, Davide, Lander, Alexander J., Li, Xuefei, Escudero, Jorge S., Tsai, Yu-Hsuan and Luk, Louis Y. P. 2020. Use of an asparaginyl endopeptidase for chemo-enzymatic peptide and protein labeling. Chemical Science 11 (23) , pp. 5881-5888. 10.1039/D0SC02023K

[img]
Preview
PDF - Published Version
Available under License Creative Commons Attribution.

Download (655kB) | Preview

Abstract

Asparaginyl endopeptidases (AEPs) are ideal for peptide and protein labeling. However, because of the reaction reversibility, a large excess of labels or backbone modified substrates are needed. In turn, simple and cheap reagents can be used to label N-terminal cysteine, but its availability inherently limits the potential applications. Aiming to address these issues, we have created a chemo-enzymatic labeling system that exploits the substrate promiscuity of AEP with the facile chemical reaction between N-terminal cysteine and 2-formyl phenylboronic acid (FPBA). In this approach, AEP is used to ligate polypeptides with a Asn–Cys–Leu recognition sequence with counterparts possessing an N-terminal Gly–Leu. Instead of being a labeling reagent, the commercially available FPBA serves as a scavenger converting the byproduct Cys–Leu into an inert thiazolidine derivative. This consequently drives the AEP labeling reaction forward to product formation with a lower ratio of label to protein substrate. By carefully screening the reaction conditions for optimal compatibility and minimal hydrolysis, conversion to the ligated product in the model reaction resulted in excellent yields. The versatility of this AEP-ligation/FPBA-coupling system was further demonstrated by site-specifically labeling the N- or C-termini of various proteins.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Chemistry
Additional Information: This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material.
Publisher: Royal Society of Chemistry
ISSN: 2041-6520
Funders: EPSRC, BBSRC, Leverhulme Trust, Royal Society and Wellcome Trust
Date of First Compliant Deposit: 16 June 2020
Date of Acceptance: 11 May 2020
Last Modified: 20 Oct 2020 17:15
URI: http://orca.cf.ac.uk/id/eprint/132485

Actions (repository staff only)

Edit Item Edit Item

Downloads

Downloads per month over past year

View more statistics