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

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Title: A cell-permeable biscyclooctyne as a novel probe for the identification of protein sulfenic acids
Authors: McGarry, DJ
Shchepinova, MM
Lilla, S
Hartley, RC
Olson, MF
Item Type: Journal Article
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 labeled proteins produced in vivo can be visualized by fluorescence microscopy to reveal their localization. These findings demonstrate a novel and multifaceted approach to the detection and trapping of sulfenic acids.
Issue Date: 28-Oct-2016
Date of Acceptance: 28-Oct-2016
URI: http://hdl.handle.net/10044/1/53437
DOI: https://dx.doi.org/10.1021/acschembio.6b00742
ISSN: 1554-8929
Publisher: American Chemical Society
Start Page: 3300
End Page: 3304
Journal / Book Title: ACS Chemical Biology
Volume: 11
Issue: 12
Copyright Statement: © 2016 American Chemical Society. ACS AuthorChoice - This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
Keywords: 03 Chemical Sciences
06 Biological Sciences
Organic Chemistry
Publication Status: Published
Open Access location: http://pubs.acs.org/doi/abs/10.1021/acschembio.6b00742
Appears in Collections:Chemistry
Biological and Biophysical Chemistry