The sulfinic acid switch in proteins

Jacob, Claus; Holme, Andrea L.; Fry, Fiona H.

doi:10.1039/b406180b

Cited by 132 publications

(81 citation statements)

References 23 publications

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“…Reversible oxidation of cysteine to disulfide (-S-S-) or sufenic acid (-SOH) is readily accomplished by thiols, such as DTT, 2-ME, glutathione, or thioredoxin. In contrast, oxidation to sulfinic acid (-SO 2 H) or sulfonic acid (-SO 3 H) is not reduced by these thiols under physiological conditions (21). For example, one cysteine in the active site of peroxiredoxin (Prx) is oxidized to sulfinic acid (-SO 2 H) by incubation with an excess of substrate of this enzyme, H 2 O 2 , and rereduced by a specific enzyme, sulfiredoxin, but not by general thiols (22,23).…”

mentioning

confidence: 98%

Oxidative Modification to Cysteine Sulfonic Acid of Cys111 in Human Copper-Zinc Superoxide Dismutase

Fujiwara

Nakano

Kato

et al. 2007

Journal of Biological Chemistry

126

142

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Copper-zinc superoxide dismutase (SOD1) plays a protective role against oxidative stress. On the other hand, recent studies suggest that SOD1 itself is a major target of oxidative damage and has its own pathogenicity in various neurodegenerative diseases, including familial amyotrophic lateral sclerosis. Only human and great ape SOD1s among mammals have the highly reactive free cysteine residue, Cys 111 , at the surface of the SOD1 molecule. The purpose of this study was to investigate the role of Cys 111 in the oxidative damage of the SOD1 protein, by comparing the oxidative susceptibility of recombinant human SOD1 modified with 2-mercaptoethanol at Cys 111 (2-ME-SOD1) to wild-type SOD1. Wild-type SOD1 was more sensitive to oxidation by hydrogen peroxide-generating fragments, oligomers, and charge isomers compared with 2-ME-SOD1. Moreover, wild-type SOD1, but not 2-ME-SOD1, generated an upper shifted band in reducing SDS-PAGE even by air oxidation. Using mass spectrometry and limited proteolysis, this upper band was identified as an oxidized subunit of SOD1; the sulfhydryl group (Cys-SH) of Cys 111 was selectively oxidized to cysteine sulfinic acid (Cys-SO 2 H) and to cysteine sulfonic acid (Cys-SO 3 H). (2). Moreover, incubation with excess H 2 O 2 caused oxidation of almost all histidine and cysteine residues (3), fragmentation (4, 5) and aggregation (6, 7) of SOD1 itself. Co-incubation with bicarbonate and H 2 O 2 also induced bicarbonate radical anion formation, resulting in oligomerization of human SOD1 (8).The familial form of amyotrophic lateral sclerosis (ALS) is associated with specific mutations in the SOD1 gene (SOD1) that encodes 153 amino acids (9, 10). To date, more than 110 familial ALS (FALS)-causing mutations in SOD1 have been identified (available on the World Wide Web); however, the mechanism by which SOD1 mutants induce ALS remains unknown. The presence of intracellular aggregates that contain SOD1 in spinal cord motor neurons is thought to be a pathological hallmark of ALS. In particular, FALS-linked mutant SOD1s are prone to misfolding and aggregation (11,12). Recently, Ezzi et al. (7) reported that even wild-type SOD1 results in aggregation after oxidation, and the oxidized wildtype SOD1 gains properties like FALS mutant SOD1s. In addition to ALS, oxidative damaged SOD1 proteins were detected in the brains of patients with Alzheimer and Parkinson diseases (13). These findings suggest that oxidized SOD1 plays a role in the pathophysiology of various neurodegenerative diseases.

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confidence: 98%

Oxidative Modification to Cysteine Sulfonic Acid of Cys111 in Human Copper-Zinc Superoxide Dismutase

Fujiwara

Nakano

Kato

et al. 2007

Journal of Biological Chemistry

126

142

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“…It is known that cysteine residues in redox regulated enzymes can be irreversibly "over-oxidized" to sulfinic and sulfonic acids by hydrogen peroxide under some conditions. 12,14,25 Two possible mechanisms can be considered for the inactivation of PTP1B by 1. Direct reaction of the active site cysteine residue with 1 would yield the inactive, sulfenic acid form of the enzyme (Scheme 2A).…”

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confidence: 99%

Oxidative inactivation of protein tyrosine phosphatase 1B by organic hydroperoxides

Bhattacharya

Labutti

Seiner

et al. 2008

Bioorganic & Medicinal Chemistry Letters

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Protein tyrosine phosphatases (PTPs) are cysteine-dependent enzymes that play a central role in cell signaling. Organic hydroperoxides cause thiol-reversible, oxidative inactivation of PTP1B in a manner that mirrors the endogenous signaling agent hydrogen peroxide.Protein tyrosine phosphatases (PTPs) are cysteine-dependent enzymes that catalyze the hydrolytic removal of phosphate groups from tyrosine residues in proteins (Scheme 1). [1][2][3] Thus, PTPs, in concert with protein tyrosine kinases, play a central role in cell signaling by regulating the phosphorylation status and, in turn, the functional properties, of target proteins in various signal transduction pathways. 4,5 The cellular activity of some PTPs is regulated by hydrogen peroxide (H 2 O 2 ) that is produced as a second messenger in response to extracellular stimuli such as insulin, epidermal growth factor, and platelet derived growth factor. 6-9 H 2 O 2 inactivates PTPs via oxidation of the active site cysteine thiol residue to a sulfenic acid. 6,10 In some cases, the cysteine sulfenic acid undergoes subsequent conversion to an active site sulfenyl amide linkage 11-13 or disulfide. 14,15 This type of oxidative inactivation of PTPs is slowly reversed upon reaction of the inactivated enzyme with biological thiols (Scheme 1). [10][11][12][13][14][15] Given the biological importance of PTPs, it may be useful to identify organic reagents that regulate PTP activity through redox mechanisms. [16][17][18][19][20] With this in mind, we examined the ability of several organic hydroperoxides (1-4) to effect thiol-reversible, oxidative inactivation of the archtypal member of the PTP family, PTP1B. 21 We find that peracetic acid (1) is a potent, time-dependent inactivator of the catalytic subunit of PTP1B (a.a. 1-322) (Fig. 1A). The observation of time-dependent inactivation is consistent with a process involving covalent modification of the enzyme. 22 A good linear fit is obtained in a plot of the apparent rates of inactivation versus concentration indicating that the inactivation reaction is a second-order process (Fig. 1B). The slope of the line reveals a rate constant of 2.1 ± 0.2 × 10 4 M −1 s −1 for the reaction of 1 with PTP1B. For comparison, hydrogen peroxide inactivates PTP1B with a rate constant of 10 M −1 s −1 . 10 The inactivation of PTP1B (250 nM) by 1 (1 μM, 3 min) is not reversed upon removal of excess inactivator by gel filtration of the enzyme through G25 Sephadex. The inactivation process is significantly slowed in the presence of phosphate ion, which is an active site-directed inhibitor of the enzyme (K i = 17 mM). Specifically, treatment of the enzyme with 1 (15 μM) for 15 s inactivates 88% of the enzyme under standard conditions, while in the presence of phosphate *Corresponding author. Tel.: +1-573-882-6763; fax: +1-573-882-2754 gatesk@missouri.edu. NIH Public Access NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript ion (50 mM) only 49% inactivation is observed. Together, the results suggest that 1 inactiv...

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“…Since many proteins rely on the unique properties of the thiol functional group for their biological activity, oxidation of specific cysteine residues can operate like a switch, activating or deactivating its cellular function in a manner analogous to more widely studied modifications, such as phosphorylation and dephosphorylation. 12,13 Despite studies implicating cysteine oxidation as a modulator of cellular processes, the molecular details of the majority of these modifications, including the complete repertoire of proteins containing thiol post-translational modifications (PTMs) and the specific sites of modification remain largely unknown. Furthermore, since thiol-modified proteins are studied in purified proteins and cell extracts the response to oxidative challenge in vitro and the importance of these modifications in a cellular context remain a hotly debated issue.…”

Section: Introductionmentioning

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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The sulfinic acid switch in proteins

Cited by 132 publications

References 23 publications

Oxidative Modification to Cysteine Sulfonic Acid of Cys111 in Human Copper-Zinc Superoxide Dismutase

Oxidative Modification to Cysteine Sulfonic Acid of Cys111 in Human Copper-Zinc Superoxide Dismutase

Oxidative inactivation of protein tyrosine phosphatase 1B by organic hydroperoxides

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

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