Thiol redox proteomics seen with fluorescent eyes: The detection of cysteine oxidative modifications by fluorescence derivatization and 2-DE

Izquierdo-Álvarez, Alicia; Martínez‐Ruiz, Antonio

doi:10.1016/j.jprot.2011.09.013

Cited by 29 publications

(14 citation statements)

References 58 publications

(142 reference statements)

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“…SMCC [succinimidyl trans ‐4‐(maleimidylmethyl)cyclohexane‐1‐carboxylate], a heterobifunctional linker containing a maleimide and an NHS ester unit] was recently used to conjugate a polyhistidine peptide to DNA capable of cross‐linking to cisplatin in order to perform “pull‐down” experiments to identify modification states of proteins binding to cisplatin . Maleimides have also been used to investigate the role of cysteine oxidation in cell signalling, for elucidating roles of endogenous cysteine residues in various protein systems, and for identifying and isolating epidermal growth factor receptor (EGFR)‐binding peptides generated by phage display …”

Section: Maleimidesmentioning

confidence: 99%

Chemical Protein Modification through Cysteine

2016

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The modification of proteins with non-protein entities is important for a wealth of applications, and methods for chemically modifying proteins attract considerable attention. Generally, modification is desired at a single site to maintain homogeneity and to minimise loss of function. Though protein modification can be achieved by targeting some natural amino acid side chains, this often leads to ill-defined and randomly modified proteins. Amongst the natural amino acids, cysteine combines advantageous properties contributing to its suitability for site-selective modification, including a unique nucleophilicity, and a low natural abundance--both allowing chemo- and regioselectivity. Native cysteine residues can be targeted, or Cys can be introduced at a desired site in a protein by means of reliable genetic engineering techniques. This review on chemical protein modification through cysteine should appeal to those interested in modifying proteins for a range of applications.

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Section: Maleimidesmentioning

confidence: 99%

Chemical Protein Modification through Cysteine

2016

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“…To identify Cys modifications, most initial redox approaches employed differential chemical or isotopic labeling followed by biotin-based affinity enrichment. In these methods, free protein thiols were initially blocked by alkylation, and then specific oxidized Cys-residues were selectively reduced and labeled with biotin tags (20), fluorophores (21,22), radioactive compounds (23), or isotope-coded affinity tags (24,25) for Western blot, gel electrophoresis, or mass spectrometry (MS) analysis. Alternatively, thiol-reactive chemical probes coupled with click chemistry have recently been applied to identify redox-sensitive proteins or Cys-residues (26,27) and in vivo thiol redox changes (28).…”

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confidence: 99%

Proteome-wide Light/Dark Modulation of Thiol Oxidation in Cyanobacteria Revealed by Quantitative Site-specific Redox Proteomics

Guo

Nguyen

Dai

et al. 2014

Molecular & Cellular Proteomics

110

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Reversible protein thiol oxidation is an essential regulatory mechanism of photosynthesis, metabolism, and gene expression in photosynthetic organisms. Herein, we present proteome-wide quantitative and site-specific profiling of in vivo thiol oxidation modulated by light/dark in the cyanobacterium Synechocystis sp. PCC 6803, an oxygenic photosynthetic prokaryote, using a resin-assisted thiol enrichment approach. Our proteomic approach integrates resin-assisted enrichment with isobaric tandem mass tag labeling to enable site-specific and quantitative measurements of reversibly oxidized thiols. The redox dynamics of ϳ2,100 Cys-sites from 1,060 proteins under light, dark, and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (a photosystem II inhibitor) conditions were quantified. In addition to relative quantification, the stoichiometry or percentage of oxidation (reversibly oxidized/total thiols) for ϳ1,350 Cys-sites was also quantified. The overall results revealed broad changes in thiol oxidation in many key biological processes, including photosynthetic electron transport, carbon fixation, and glycolysis. Moreover,

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“…Protein S-nitrosylation can be detected by specific fluorescent labelling with a “fluorescence switch” procedure [48, 51]. We therefore used this technique to evaluate the extent of S-nitrosylation in erythrocytes (Fig.…”

Section: Resultsmentioning

confidence: 99%

Trifluoperazine-Induced Suicidal Erythrocyte Death and S-Nitrosylation Inhibition, Reversed by the Nitric Oxide Donor Sodium Nitroprusside

Ghashghaeinia

Wesseling

Ramos

et al. 2017

Cell Physiol Biochem

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Background and Purpose: The high potency antipsychotic drug trifluoperazine (10-[3-(4-methyl-1-piperazinyl)-propyl]-2-(trifluoromethyl)-(10)H-phenothiazine dihydrochloride; TFP) may either counteract or promote suicidal cell death or apoptosis. Similar to apoptosis, erythrocytes may enter eryptosis, characterized by phosphatidylserine exposure at the cell surface and cell shrinkage. Eryptosis can be stimulated by an increase in cytoplasmic Ca²⁺ concentration ([Ca²⁺]_i) and inhibited by nitric oxide (NO). We explored whether TFP treatment of erythrocytes induces phosphatidylserine exposure, cell shrinkage, and calcium influx, whether it impairs S-nitrosylation and whether these effects are inhibited by NO. Methods: Phosphatidylserine exposure at the cell surface was estimated from annexin-V-binding, cell volume from forward scatter, [Ca²⁺]_i from Fluo3-fluorescence, and protein nitrosylation from fluorescence switch of the Bodipy-TMR/Sypro Ruby signal. Results: Exposure of human erythrocytes to TFP significantly enhanced the percentage of annexin-V-binding cells, raised [Ca²⁺]_i, and decreased S-nitrosylation. The effect of TFP on annexin-V-binding was not affected by removal of extracellular Ca²⁺ alone, but was significantly inhibited by pre-treatment with sodium nitroprusside (SNP), an effect significantly augmented by additional removal of extracellular Ca²⁺. A 3 hours treatment with 0.1 µM Ca²⁺ ionophore ionomycin triggered annexin-V-binding and cell shrinkage, effects fully reversed by removal of extracellular Ca²⁺. Conclusions: TFP induces eryptosis and decreases protein S-nitrosylation, effects blunted by nitroprusside. The effect of nitroprusside is attenuated in the presence of extracellular Ca²⁺.

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Thiol redox proteomics seen with fluorescent eyes: The detection of cysteine oxidative modifications by fluorescence derivatization and 2-DE

Cited by 29 publications

References 58 publications

Chemical Protein Modification through Cysteine

Chemical Protein Modification through Cysteine

Proteome-wide Light/Dark Modulation of Thiol Oxidation in Cyanobacteria Revealed by Quantitative Site-specific Redox Proteomics

Trifluoperazine-Induced Suicidal Erythrocyte Death and S-Nitrosylation Inhibition, Reversed by the Nitric Oxide Donor Sodium Nitroprusside

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