Mass spectrometry in studies of protein thiol chemistry and signaling: Opportunities and caveats

Baez, Nelmi O. Devarie; Reisz, Julie A.; Furdui, Cristina M.

doi:10.1016/j.freeradbiomed.2014.09.016

Cited by 56 publications

(29 citation statements)

References 206 publications

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“…Indeed, many proteomic approaches aimed at studying disulfide pairs in proteins require considerable sample work up ex vivo, often employing a step to block free thiols and, therefore, prevent undesirable disulfide rearrangements [7]. In addition, the limited bioavailability, bulkiness, and slow reactivity of many biorthogonal oxidation traps that react with cysteine sulfenic acids pose challenges in identifying oxidation-prone proteins in cells [30, 66, 67]. …”

Section: Discussionmentioning

confidence: 99%

Trapping redox partnerships in oxidant-sensitive proteins with a small, thiol-reactive cross-linker

Allan

Loberg

Chepngeno

et al. 2016

Free Radical Biology and Medicine

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A broad range of redox-regulated proteins undergo reversible disulfide bond formation on oxidation-prone cysteine residues. Heightened reactivity of the thiol groups in these cysteines also increases susceptibility to modification by organic electrophiles, a property that can be exploited in the study of redox networks. Here, we explored whether divinyl sulfone (DVSF), a thiol-reactive bifunctional electrophile, cross-links oxidant-sensitive proteins to their putative redox partners in cells. To test this idea, previously identified oxidant targets involved in oxidant defense (namely, peroxiredoxins, methionine sulfoxide reductases, sulfiredoxin, and glutathione peroxidases), metabolism, and proteostasis were monitored for cross-link formation following treatment of Saccharomyces cerevisiae with DVSF. Several proteins screened, including multiple oxidant defense proteins, underwent intermolecular and/or intramolecular cross-linking in response to DVSF. Specific redox-active cysteines within a subset of DVSF targets were found to influence cross-linking; in addition, DVSF-mediated cross-linking of its targets was impaired in cells first exposed to oxidants. Since cross-linking appeared to involve redox-active cysteines in these proteins, we examined whether potential redox partners became cross-linked to them upon DVSF treatment. Specifically, we found that several substrates of thioredoxins were cross-linked to the cytosolic thioredoxin Trx2 in cells treated with DVSF. However, other DVSF targets, like the peroxiredoxin Ahp1, principally formed intra-protein cross-links upon DVSF treatment. Moreover, additional protein targets, including several known to undergo S-glutathionylation, were conjugated via DVSF to glutathione. Our results indicate that DVSF is of potential use as a chemical tool for irreversibly trapping and discovering thiol-based redox partnerships within cells.

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Section: Discussionmentioning

confidence: 99%

Trapping redox partnerships in oxidant-sensitive proteins with a small, thiol-reactive cross-linker

Allan

Loberg

Chepngeno

et al. 2016

Free Radical Biology and Medicine

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“…This delicate balance of productive versus pathogenic reactions occurring among thiol groups presents researchers with the challenge of identifying not only the specific reaction products and sites of modification occurring on thiol groups of small molecules and proteins, but also the functional effect of each type and location of modification. Fortunately, the tools and approaches for evaluating this complex array of reactivities and products have been improving over the last decade and are the subject of other reviews in this series [24] and elsewhere [25–28]. …”

Section: Reactions Of Cysteines and Other Thiol-containing Biomoleculesmentioning

confidence: 99%

The basics of thiols and cysteines in redox biology and chemistry

Poole

2015

Free Radical Biology and Medicine

804

679

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Cysteine is one of the least abundant amino acids, yet it is frequently found as a highly conserved residue within functional (regulatory, catalytic or binding) sites in proteins. It is the unique chemistry of the thiol or thiolate group of cysteine that imparts functional sites with their specialized properties (e.g., nucleophilicity, high affinity metal binding, and/or ability to form disulfide bonds). Highlighted in this review are some of the basic biophysical and biochemical properties of cysteine groups and the equations that apply to them, particularly with respect to pKa and redox potential. Also summarized are the types of low molecular weight thiols present in high concentrations in most cells, as well as the ways in which modifications of cysteinyl residues can impart or regulate molecular functions important to cellular processes including signal transduction.

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“…These rely on either the derivatization of -SOH to a stable conjugate with the use of selective chemical probes or tag-switch methods of analysis consisting of a blocking step, selective -SOH reduction to -SH state, and reaction of newly formed thiols with a distinct chemical reagent (reviewed in [30]). Direct chemical derivatization of -SOH is possible by exploiting both the electrophilicity and nucleophilicity of the sulfenyl moiety.…”

Section: Synthesis Stability Reactivity and Characterization Of mentioning

confidence: 99%

Biological chemistry and functionality of protein sulfenic acids and related thiol modifications

Devarie‐Baez

Lopez

Furdui

2015

Free Radical Research

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Selective modification of proteins at cysteine residues by reactive oxygen, nitrogen or sulfur species formed under physiological and pathological states is emerging as a critical regulator of protein activity impacting cellular function. This review focuses primarily on protein sulfenylation (-SOH), a metastable reversible modification connecting reduced cysteine thiols to many products of cysteine oxidation. An overview is first provided on the chemistry principles underlining synthesis, stability and reactivity of sulfenic acids in model compounds and proteins, followed by a brief description of analytical methods currently employed to characterize these oxidative species. The following chapters present a selection of redox-regulated proteins for which the -SOH formation was experimentally confirmed and linked to protein function. These chapters are organized based on the participation of these proteins in the regulation of signaling, metabolism and epigenetics. The last chapter discusses the therapeutic implications of altered redox microenvironment and protein oxidation in disease.

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Mass spectrometry in studies of protein thiol chemistry and signaling: Opportunities and caveats

Cited by 56 publications

References 206 publications

Trapping redox partnerships in oxidant-sensitive proteins with a small, thiol-reactive cross-linker

Trapping redox partnerships in oxidant-sensitive proteins with a small, thiol-reactive cross-linker

The basics of thiols and cysteines in redox biology and chemistry

Biological chemistry and functionality of protein sulfenic acids and related thiol modifications

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