A cell-permeable biscyclooctyne as a novel probe for the identification of protein sulfenic acids

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)
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 > Institute 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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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
682111Cysteine oxidation as a regulator of cancer cell motility and invasionMichael OlsonMedical Research Council (MRC)MR/M018776/1ICS - BEATSON INSTITUTE FOR CANCER RES.
694461EPSRC Centre for Multiscale soft tissue mechanics with application to heart & cancerRaymond OgdenEngineering & Physical Sciences Research Council (EPSRC)EP/N014642/1M&S - MATHEMATICS
584861EPSRC Doctoral Training Grant 2011-2015Mary Beth KneafseyEngineering & Physical Sciences Research Council (EPSRC)EP/J500434/1VPO VICE PRINCIPAL RESEARCH & ENTERPRISE
584862EPSRC Doctoral Training Grant 2011-2015Mary Beth KneafseyEngineering & Physical Sciences Research Council (EPSRC)EP/J500434/1VPO VICE PRINCIPAL RESEARCH & ENTERPRISE