Dual-Reactivity trans-Cyclooctenol Probes for Sulfenylation in Live Cells Enable Temporal Control via Bioorthogonal Quenching

Scinto, Samuel L.; Ekanayake, Oshini; Seneviratne, Uthpala; Pigga, Jessica E.; Boyd, Samantha J.; Taylor, Michael T.; Li, Jun; Ende, Christopher W. am; Rozovsky, Sharon; Fox, Joseph M.

doi:10.1021/jacs.9b01164

Cited by 41 publications

(38 citation statements)

References 48 publications

(93 reference statements)

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“…A total of 148 out of 473 proteins were detected that had not been previously identified in other surveys of the sulfenome. This result reveals that the norbornene probes for cysteine sulfenic acid are complementary to previously reported probes, exhibiting excellent chemoselectivity and a distinct coverage of the sulfenome network. The full list of protein hits identified by using the norbornene probe provides motivation for further analysis of the role of cysteine oxidation in cellular function and disease.…”

Section: Figurementioning

confidence: 99%

Proteome‐Wide Survey of Cysteine Oxidation by Using a Norbornene Probe

et al. 2020

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Rapid detection of cysteine oxidation in living cells is critical in advancing our understanding of responses to reactive oxygen species (ROS) and oxidative stress. Accordingly, there is a need to develop chemical probes that facilitate proteome‐wide detection of cysteine's many oxidation states. Herein, we report the first whole‐cell proteomics analysis using a norbornene probe to detect the initial product of cysteine oxidation: cysteine sulfenic acid. The oxidised proteins identified in the HeLa cell model represent the first targets of the ROS hydrogen peroxide. The panel of protein hits provides new and important information about the targets of oxidative stress, including 148 new protein members of the sulfenome. These findings provide new leads for the study and understanding of redox signalling and diseases associated with oxidative stress.

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Section: Figurementioning

confidence: 99%

Proteome‐Wide Survey of Cysteine Oxidation by Using a Norbornene Probe

et al. 2020

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“…This result reveals that the norbornene probes for cysteine sulfenic acid are complementary to dimedone and related nucleophilic probes. 45 The full list of protein hits identified using the norbornene probe provides motivation for further analysis into the role of cysteine oxidation in cellular function and disease. Our own laboratories are pursing these hits further with an aim to discover new roles for cysteine oxidation.…”

Section: Resultsmentioning

confidence: 99%

Proteome-Wide Survey Reveals Norbornene Is Complementary to Dimedone and Related Nucleophilic Probes for Cysteine Sulfenic Acid

Alcock¹,

Langini²,

Remke³

et al. 2019

Preprint

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Detection of cysteine sulfenic acid in live cells is critical in advancing our understanding of cysteine redox chemistry and its biological function. Accordingly, there is a need to develop sulfenic acid-specific chemical probes with distinct reaction mechanisms to facilitate proteome-wide detection of this important posttranslational modification. Herein, we report the first whole-cell proteomics analysis using a norbornene probe to detect cysteine sulfenic acid in live HeLa cells. Comparison of the enriched proteins to those identified using dimedone and other C-nucleophilic probes revealed a complementary reactivity profile. Remarkably, 148 new members of the sulfenome were identified. These discoveries highlight how subtle differences in chemical reactivity of both the probes and cysteine residues influence detection. Overall, this study expands our understanding of protein oxidation at cysteine and reveals new proteins to consider for future studies of cysteine oxidation, redox regulation and signaling, and the biochemistry of oxidative stress.

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“…The expanded palette of carbon nucleophiles with enhanced SOH reactivity [161], set the stage to develop novel clickable tools to assess SOH using Click PEGylation. Furthermore, Fox' group [162] have developed SOH reactive TCO, which enables Tz quenching to avoid ex vivo artefacts; such tools could be modified to enable IEDDA mediated Click PEGylation. Novel DiaAlk reagents afford similar opportunities to selectively assess SO 2 [147].…”

Section: Solutionsmentioning

confidence: 99%

Immunological Techniques to Assess Protein Thiol Redox State: Opportunities, Challenges and Solutions

Cobley

Husi

2020

Antioxidants

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To understand oxidative stress, antioxidant defense, and redox signaling in health and disease it is essential to assess protein thiol redox state. Protein thiol redox state is seldom assessed immunologically because of the inability to distinguish reduced and reversibly oxidized thiols by Western blotting. An underappreciated opportunity exists to use Click PEGylation to realize the transformative power of simple, time and cost-efficient immunological techniques. Click PEGylation harnesses selective, bio-orthogonal Click chemistry to separate reduced and reversibly oxidized thiols by selectively ligating a low molecular weight polyethylene glycol moiety to the redox state of interest. The resultant ability to disambiguate reduced and reversibly oxidized species by Western blotting enables Click PEGylation to assess protein thiol redox state. In the present review, to enable investigators to effectively harness immunological techniques to assess protein thiol redox state we critique the chemistry, promise and challenges of Click PEGylation.

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Dual-Reactivity trans-Cyclooctenol Probes for Sulfenylation in Live Cells Enable Temporal Control via Bioorthogonal Quenching

Cited by 41 publications

References 48 publications

Proteome‐Wide Survey of Cysteine Oxidation by Using a Norbornene Probe

Proteome‐Wide Survey of Cysteine Oxidation by Using a Norbornene Probe

Proteome-Wide Survey Reveals Norbornene Is Complementary to Dimedone and Related Nucleophilic Probes for Cysteine Sulfenic Acid

Immunological Techniques to Assess Protein Thiol Redox State: Opportunities, Challenges and Solutions

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