Reactivity, Selectivity, and Stability in Sulfenic Acid Detection: A Comparative Study of Nucleophilic and Electrophilic Probes

Gupta, Vinayak; Paritala, Hanumantharao; Carroll, Kate S.

doi:10.1021/acs.bioconjchem.6b00181

Cited by 51 publications

(57 citation statements)

References 59 publications

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“…S1) yielded similar levels of d 6 -dimedone labeling, which may be due in part to the one hour incubation time with microsomes. While dimedone has been shown to be selective, the low reactivity of dimedone with sulfenic acids requires extended incubation times to achieve sufficient labeling (rate of 0.8 min -1 under the reported conditions) (39,40). The modified cysteines identified are likely sensitive to oxidation, but the amount of labeling observed may be representative of the equilibrium of a saturated system.…”

Section: Discussionmentioning

confidence: 93%

Sulfenylation of Human Liver and Kidney Microsomal Cytochromes P450 and Other Drug-Metabolizing Enzymes as a Response to Redox Alteration

Albertolle

Phan

Pozzi

et al. 2018

Molecular & Cellular Proteomics

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2The abbreviations used are: CID, collision-induced dissociation; DTT, dithiothreitol; ER, endoplasmic reticulum; FDR, (peptide) false discovery rate; FMO, microsomal flavin-containing monooxygenase; HCD, higher energy collisional dissociation; HEPPS, 3-[4-(2-hydroxyethyl)-1-piperazinyl]propanesulfonate; ICDID, isotope-coded dimedone/iododimedone (labeling); MAO, monoamine oxidase; P450 or CYP, cytochrome P450; TCEP, tris(carbethoxymethyl)phosphine;UGT, UDP-glucuronyl transferase. SUMMARYThe lumen of the endoplasmic reticulum (ER) provides an oxidizing environment to aid in the formation of disulfide bonds, which is tightly regulated by both antioxidant proteins and small molecules. On the cytoplasmic side of the ER, cytochrome P450 (P450) proteins have been identified as a superfamily of enzymes that are important in the formation of endogenous chemicals as well as in the detoxication of xenobiotics. Our previous report described oxidative inhibition of P450 Family 4 enzymes via oxidation of the heme-thiolate cysteine to a sulfenic acid (-SOH) (Albertolle, M. E. et al. (2017) J. Biol. Chem. 292, 11230-11242). Further proteomic analyses of murine kidney and liver microsomes led to the finding that a number of other drug metabolizing enzymes located in the ER are also redox-regulated in this manner. We expanded our analysis of sulfenylated enzymes to human liver and kidney microsomes. Evaluation of the sulfenylation, catalytic activity, and spectral properties of P450s 1A2, 2C8, 2D6, and 3A4 led to the identification of two classes of redox sensitivity in P450 enzymes: heme thiolate-sensitive and thiol-insensitive. These findings provide evidence for a mammalian P450 regulatory mechanism, which may also be relevant to other drug-metabolizing enzymes. (Data are available via ProteomeXchange with identifier PXD007913.) (INTRODUCTION)The endoplasmic reticulum (ER) is a site of protein translation, posttranslational processing, and small molecule metabolism (1). Posttranslational processing includes glycosylation of proteins for sorting, disulfide bond formation, and specific proteolytic cleavages.The ER lumen is an oxidizing environment that aids in the production of disulfide bonds, which is maintained at a homeostatic level by both small molecules (ascorbate) and proteins (protein disulfide isomerases) (2, 3). The cytosolic side of the ER remains a reducing environment to preserve the normal function of integral ER proteins.Cytochromes P450 (CYP, P450) are found in the cytoplasmic side of the ER and are most well-known for the ability to metabolize xenobiotics and important endogenous substrates (e.g., steroids and vitamins) (4). These proteins have been of interest in the pharmaceutical industry since their initial discovery, and enzymes in P450 Subfamilies 1A, 2C, 2D, and 3A are involved in the metabolism and clearance of a large majority of small molecule drugs currently approved for clinical use in humans (4). P450 regulation involves genetic and epigenetic aspects, as well as transcriptional regulation by bot...

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Section: Discussionmentioning

confidence: 93%

Sulfenylation of Human Liver and Kidney Microsomal Cytochromes P450 and Other Drug-Metabolizing Enzymes as a Response to Redox Alteration

Albertolle

Phan

Pozzi

et al. 2018

Molecular & Cellular Proteomics

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“…4A could be also due to protein sulfenation which is a reversible process [71–73]. Mitochondrial proteins were labeled with a specific sulfenation probe DCP-Bio1 [74] followed by BN-PAGE isolation of DLDH and Western blot detection of DLDH sulfenation.…”

Section: Resultsmentioning

confidence: 99%

Administration of 5-methoxyindole-2-carboxylic acid that potentially targets mitochondrial dihydrolipoamide dehydrogenase confers cerebral preconditioning against ischemic stroke injury

et al. 2017

Free Radical Biology and Medicine

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The objective of this study was to investigate a possible role of mitochondrial dihydrolipoamide dehydrogenase (DLDH) as a chemical preconditioning target for neuroprotection against ischemic injury. We used 5-methoxyindole-2-carboxylic acid (MICA), a reportedly reversible DLDH inhibitor, as the preconditioning agent and administered MICA to rats mainly via dietary intake. Upon completion of 4 week’s MICA treatment, rats underwent 1 h transient ischemia and 24 h reperfusion followed by tissue collection. Our results show that MICA protected the brain against ischemic stroke injury as the infarction volume of the brain from the MICA-treated group was significantly smaller than that from the control group. Data were then collected without or with stroke surgery following MICA feeding. It was found that in the absence of stroke following MICA feeding, DLDH activity was lower in the MICA treated group than in the control group, and this decreased activity could be partly due to DLDH protein sulfenation. Moreover, DLDH inhibition by MICA was also found to upregulate the expression of NAD(P)H-ubiquinone oxidoreductase 1(NQO1) via the Nrf2 signaling pathway. In the presence of stroke following MICA feeding, decreased DLDH activity and increased Nrf2 signaling were also observed along with increased NQO1 activity, decreased oxidative stress, decreased cell death, and increased mitochondrial ATP output. We also found that MICA had a delayed preconditioning effect four weeks post MICA treatment. Our study indicates that administration of MICA confers chemical preconditioning and neuroprotection against ischemic stroke injury.

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“…A strongly positive aspect of using DCP-Bio1 to detect protein oxidation is that it labels cysteines undergoing direct and active oxidation. However, it should be noted that, because DCP-Bio1 detects only the likely transient sulfenic acid (or potentially sulfenylamide [16, 17]) intermediate, it does not report on the total pool of oxidized protein.…”

Section: Discussionmentioning

confidence: 99%

“…Much work has been done by us and others in the field of redox signaling to identify targets of signal-generated ROS [14–17]. Various ROS, reactive nitrogen species (RNS), and reactive sulfur species (RSS) react with cysteine residues of proteins, modifying the structure, dynamics, and/or function of those proteins.…”

mentioning

confidence: 99%

Endogenous, regulatory cysteine sulfenylation of ERK kinases in response to proliferative signals

Keyes

Parsonage

Yammani

et al. 2017

Free Radical Biology and Medicine

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ERK-dependent signaling is key to many pathways through which extracellular signals are transduced into cell-fate decisions. One conundrum is the way in which disparate signals induce specific responses through a common, ERK-dependent kinase cascade. While studies have revealed intricate ways of controlling ERK signaling through spatiotemporal localization and phosphorylation dynamics, additional modes of ERK regulation undoubtedly remain to be discovered. We hypothesized that fine-tuning of ERK signaling could occur by cysteine oxidation. We report that ERK is actively and directly oxidized by signal-generated H2O2 during proliferative signaling, and that ERK oxidation occurs downstream of a variety of receptor classes tested in four cell lines. Furthermore, within the tested cell lines and proliferative signals, we observed that both activation loop-phosphorylated and non-phosphorylated ERK undergo sulfenylation in cells and that dynamics of ERK sulfenylation is dependent on the cell growth conditions prior to stimulation. We also tested the effect of endogenous ERK oxidation on kinase activity and report that phosphotransfer reactions are reversibly inhibited by oxidation by as much as 80 to 90%, underscoring the importance of considering this additional modification when assessing ERK activation in response to extracellular signals.

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Reactivity, Selectivity, and Stability in Sulfenic Acid Detection: A Comparative Study of Nucleophilic and Electrophilic Probes

Cited by 51 publications

References 59 publications

Sulfenylation of Human Liver and Kidney Microsomal Cytochromes P450 and Other Drug-Metabolizing Enzymes as a Response to Redox Alteration

Sulfenylation of Human Liver and Kidney Microsomal Cytochromes P450 and Other Drug-Metabolizing Enzymes as a Response to Redox Alteration

Administration of 5-methoxyindole-2-carboxylic acid that potentially targets mitochondrial dihydrolipoamide dehydrogenase confers cerebral preconditioning against ischemic stroke injury

Endogenous, regulatory cysteine sulfenylation of ERK kinases in response to proliferative signals

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