McGarry, D. J., Shchepinova, M. M., Lillla, S., Hartley, R. C. and Olson, M. F. (2016) A cell-permeable biscyclooctyne as a novel probe for the identification of protein sulfenic acids. ACS Chemical Biology, 11(12), pp. 3300-3304. (doi: 10.1021/acschembio.6b00742) (PMID:27792307)
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Abstract
Reactive oxygen species act as important second messengers in cell signaling and homeostasis through the oxidation of protein thiols. However, the dynamic nature of protein oxidation and the lack of sensitivity of existing molecular probes have hindered our understanding of such reactions; therefore, new tools are required to address these challenges. We designed a bifunctional variant of the strained bicyclo[6.1.0]nonyne (BCN-E-BCN) that enables the tagging of intracellular protein sulfenic acids for biorthogonal copper-free click chemistry. In validation studies, BCN-E-BCN binds the sulfenylated form of the actin-severing protein cofilin, while mutation of the cognate cysteine residues abrogates its binding. BCN-E-BCN is cell permeable and reacts rapidly with cysteine sulfenic acids in cultured cells. Using different azide-tagged conjugates, we demonstrate that BCN-E-BCN can be used in various applications for the detection of sulfenylated proteins. Remarkably, cycloaddition of an azide-tagged fluorophore to BCN-E-BCN labelled proteins produced in vivo can be visualized by fluorescence microscopy to reveal their subcellular localization. These findings demonstrate a novel and multifaceted approach to the detection and trapping of sulfenic acids.
Item Type: | Articles (Letter) |
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Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Hartley, Professor Richard and Olson, Professor Michael |
Authors: | McGarry, D. J., Shchepinova, M. M., Lillla, S., Hartley, R. C., and Olson, M. F. |
College/School: | College of Medical Veterinary and Life Sciences > School of Cancer Sciences College of Science and Engineering > School of Chemistry |
Journal Name: | ACS Chemical Biology |
Publisher: | American Chemical Society |
ISSN: | 1554-8929 |
ISSN (Online): | 1554-8937 |
Published Online: | 28 October 2016 |
Copyright Holders: | Copyright © 2016 American Chemical Society |
First Published: | First published in ACS Chemistry 11(12):3300–3304 |
Publisher Policy: | Reproduced under a creative commons license |
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