Protein Sulfenation as a Redox Sensor

Charles, Rebecca L.; Schröder, Ewald; May, Georgina; Free, Paul; Gaffney, Piers R. J.; Wait, Robin; Begum, Shajna; Heads, Richard J.; Eaton, Philip

doi:10.1074/mcp.m700065-mcp200

Cited by 175 publications

(72 citation statements)

References 48 publications

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“…Moreover, once introduced into proteins the azide group can be tagged by a wide variety of affinity and fluorescent reagents ex vivo. 33 While the synthesis of 1,3-cyclohexanedione analogs linked to fluorophores or biotin has been reported, 27,30,31 the specificity of these reagents for sulfenic acids has not been rigorously evaluated. In order to test the effects of functionalizing 1,3-cyclohexanedione on its overall reactivity, two model proteins were examined in vitro.…”

Section: Resultsmentioning

confidence: 99%

A chemical approach for detecting sulfenic acid-modified proteins in living cells

et al. 2008

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Oxidation of the thiol functional group in cysteine (Cys-SH) to sulfenic (Cys-SOH), sulfinic (Cys-SO 2 H) and sulfonic acids (Cys-SO 3 H) is emerging as an important post-translational modification that can activate or deactivate the function of many proteins. Changes in thiol oxidation state have been implicated in a wide variety of cellular processes and correlate with disease states but are difficult to monitor in a physiological setting because of a lack of experimental tools. Here, we describe a method that enables live cell labeling of sulfenic acidmodified proteins. For this approach, we have synthesized the probe DAz-1, which is chemically selective for sulfenic acids and cell permeable. In addition, DAz-1 contains an azide chemical handle that can be selectively detected with phosphine reagents via the Staudinger ligation for identification, enrichment and visualization of modified proteins. Through a combination of biochemical, mass spectrometry and immunoblot approaches we characterize the reactivity of DAz-1 and highlight its utility for detecting protein sulfenic acids directly in mammalian cells. This novel method to isolate and identify sulfenic acid-modified proteins should be of widespread utility for elucidating signaling pathways and regulatory mechanisms that involve oxidation of cysteine residues.

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Section: Resultsmentioning

confidence: 99%

A chemical approach for detecting sulfenic acid-modified proteins in living cells

et al. 2008

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“…12D) [149]. Biotin-dimedone incorporation was monitored by confocal fluorescence microscopy after staining permeabilized cells with ExtrAvidin-FITC.…”

Section: Sulfenic Acids In Proteinsmentioning

confidence: 99%

Sulfenic acid chemistry, detection and cellular lifetime

Gupta

Carroll

2014

Biochimica et Biophysica Acta (BBA) - General Subjects

371

399

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Background Reactive oxygen species-mediated cysteine sulfenic acid modification has emerged as an important regulatory mechanism in cell signaling. The stability of sulfenic acid in proteins is dictated by the local microenvironment and ability of antioxidants to reduce this modification. Several techniques for detecting this cysteine modification have been developed, including direct and in situ methods. Scope of review This review presents a historical discussion of sulfenic acid chemistry and highlights key examples of this modification in proteins. A comprehensive survey of available detection techniques with advantages and limitations is discussed. Finally, issues pertaining to rates of sulfenic acid formation, reduction, and chemical trapping methods are also covered. Major conclusions Early chemical models of sulfenic acid yielded important insights into the unique reactivity of this species. Subsequent pioneering studies led to the characterization of sulfenic acid formation in proteins. In parallel, the discovery of oxidant-mediated cell signaling pathways and pathological oxidative stress has led to significant interest in methods to detect these modifications. Advanced methods allow for direct chemical trapping of protein sulfenic acids directly in cells and tissues. At the same time, many sulfenic acids are short-lived and the reactivity of current probes must be improved to sample these species, while at the same time, preserving their chemical selectivity. Inhibitors with binding scaffolds can be rationally designed to target sulfenic acid modifications in specific proteins. General significance Ever increasing roles for protein sulfenic acids have been uncovered in physiology and pathology. A more complete understanding of sulfenic acid-mediated regulatory mechanisms will continue to require rigorous and new chemical insights. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.

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“…The ␤ globin is characterized by several oxidizable residues, such as 2 Cys and one Met residue. To understand whether the oxygen addition is due to sulfenic acid, the reaction mixture was incubated with the nucleophilic reagent dimedone (5,5-dimethyl-1,3-cyclohexanedione), which reacts specifically with sulfenic acid to form a stable thioether adduct [20]. As shown in Fig.…”

Section: Figurementioning

confidence: 99%