Strategy for identification and detection of multiple oxidative modifications within proteins applied on persulfate‐oxidized hemoglobin and human serum albumin

Mörtstedt, Harriet; Jeppsson, Marina C.; Ferrari, Giovanni; Jönsson, Bo; Kåredal, Monica; Lindh, Christian

doi:10.1002/rcm.4867

Cited by 7 publications

(7 citation statements)

References 30 publications

(39 reference statements)

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“…It has previously been shown that persulfates oxidize albumin in vitro . In these studies, the amino acids tryptophan and methionine were found to be oxidized. , Therefore, we wanted to measure oxidized peptides in these persulfate-challenged subjects to evaluate whether these oxidized peptides could be used as biomarkers for persulfate exposure or effect, for example, oxidative stress. We presented a simple SRM-based method for quantification of multiple oxidations without the use of any prior enrichment.…”

Section: Discussionmentioning

confidence: 99%

“…SRM assays targeting oxidized peptides were developed using Skyline (Version 1.4.0) . NLF proteins and peptides that previously had been shown to be targets for oxidation were selected from the scientific literature. ,, Both the modified and the corresponding unmodified tryptic peptide were included. Development of SRM assays targeting oxidized peptides is described in the Supporting Information.…”

Section: Methodsmentioning

confidence: 99%

“…7 The direct action of persulfate salts on cells and biomolecules has also been proposed as a possible mechanism. Persulfates are oxidizing agents, and they have been shown to oxidize proteins in vitro 12,13 and induce generation of reactive oxygen species (ROS) from mast cells and basophils. 13 In the present study, 159 nasal lavage fluid (NLF) samples from persulfate-challenged subjects were analyzed with liquid chromatography−tandem mass spectrometry using selected reaction monitoring (LC-SRM-MS) targeting 246 NLF proteins.…”

Section: ■ Introductionmentioning

confidence: 99%

“…The direct action of persulfate salts on cells and biomolecules has also been proposed as a possible mechanism. Persulfates are oxidizing agents, and they have been shown to oxidize proteins in vitro , and induce generation of reactive oxygen species (ROS) from mast cells and basophils …”

Section: Introductionmentioning

confidence: 99%

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Targeted Proteomic Analyses of Nasal Lavage Fluid in Persulfate-Challenged Hairdressers with Bleaching Powder-Associated Rhinitis

et al. 2015

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Hairdressers have an increased risk for developing airway symptoms, for example, asthma and rhinitis. Persulfates, which are oxidizing agents in bleaching powder, are considered important causal agents for these symptoms. However, the underlying mechanisms are unclear. The aim was therefore to measure proteomic changes in nasal lavage fluid from persulfate-challenged subjects to identify proteins potentially involved in the pathogenesis of bleaching powder-associated rhinitis or candidate effect biomarkers for persulfate. Also, oxidized peptides were measured to evaluate their usefulness as biomarkers for persulfate exposure or effect, for example, oxidative stress. Samples from hairdressers with and without bleaching powder-associated rhinitis were analyzed with liquid chromatography tandem mass spectrometry using selected reaction monitoring to target 246 proteins and five oxidized peptides. Pathway analysis was applied to obtain a functional overview of the proteins. Several proteins involved in biologically meaningful pathways, functions, or disorders, for example, inflammatory responses, oxidative stress, epithelium integrity, and dermatological disorders, changed after the persulfate challenge. A list with nine proteins that appeared to be affected by the persulfate challenge and should be followed up was defined. An albumin peptide containing oxidized tryptophan increased 2 h and 5 h after the challenge but not after 20 min, which indicates that such peptides may be useful as oxidative stress biomarkers.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Targeted Proteomic Analyses of Nasal Lavage Fluid in Persulfate-Challenged Hairdressers with Bleaching Powder-Associated Rhinitis

et al. 2015

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“…protein within the gel band may be achieved (Figures 2(b) and 3). Most studies have concentrated on the study of human albumin, [31][32][33][34] but the works by Maisonneuve et al 35 and Guedes et al 19 provide a detailed description of BSA oxidation under MCO conditions. Maisonneuve et al have described the identification of seven lysine and four arginine residues as targets for MCO.…”

Section: Protein Identification and Semi-quantification By Nano-rp-hpmentioning

confidence: 99%

Efficiency of Trypsin Digestion for Mass-Spectrometry-Based Identification and Quantification of Oxidized Proteins: Evaluation of the Digestion of Oxidized Bovine Serum Albumin

Gouveia

Silva

Vitorino

et al. 2014

Eur J Mass Spectrom (Chichester)

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In bottom-up proteomics approaches, the enzymatic proteolysis step before mass spectrometry (MS) analysis is of crucial importance, as only the efficient digestion of the protein will ensure its accurate quantification. The structural and chemical alterations occurring upon protein oxidation may decrease the efficiency of trypsin digestion, compromising the ensuing MS analysis. Herein, the efficiency of the trypsin digestion of oxidized bovine serum albumin (BSA) was assessed by protein-sequence coverage and the exponentially modified protein abundance index (emPAI) algorithm, allowing a comparison of protein abundance in samples with different levels of oxidation. Despite the extensive oxidation induced to BSA, verified by analysis of protein carbonyls, no significant difference in the yield of tryptic peptides from oxidized samples could be observed by nano-high-performance liquid chromatography (HPLC) and nano-HPLC7-electrospray ionization-MS analysis. After a database search, similar protein-sequence coverage rates were obtained for both treated and control samples. Thus, exponentially modified protein abundance index scores confirmed that, regardless of being oxidized, the same amount of BSA was present in the sodium dodecyl sulfate/polyacrylamide gel electrophoresis bands excised for digestion. The obtained results show that the digestion of the control and oxidized samples were similar, leading to the conclusion that in-gel proteolysis is not a main hindrance for the identification and quantification of oxidized proteins by MS.

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β‐cyclodextrin encapsulated polyphenols as effective antioxidants

et al. 2017

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Formation of dityrosine (DT) cross-linkages in proteins is one of the most widely used markers of oxidative stress. Ribonuclease A (RNase A) has 6 Tyr residues and shows a characteristic DT fluorescence peak upon oxidation in addition to major changes in its secondary structure. DT formation can be prevented by using polyphenols (GA, ECG, and EGCG) which are known to have strong antioxidant activity. However, it has been observed that ECG and EGCG initiate protein oligomerization due to protein-polyphenol cross-linkages. To prevent the formation of such crosslinkages we have used b-cyclodextrin (b-CD) to encapsulate the polyphenols and studied its antioxidant properties along with that of free polyphenols. The polyphenol/b-cyclodextrin (b-CD) inclusion complexes not only prevent DT formation but also reduce protein oligomerization. This may be attributed to the fact that the quinone forming rings of ECG and EGCG become encapsulated in the cavity of b-CD and are no longer available for protein cross-linking.b-cyclodextrin encapsulation, dityrosine, oxidative stress, polyphenols, protein oligomerization | I N TR ODU C TI ONBiological aging in mammalian cells is a direct indication of oxidative stress in proteins. [1] These oxidatively modified forms of proteins are generated due to the presence of reactive oxygen species (ROS) such as hydroxyl and superoxide free radicals. [2,3] Persulfates, a key ingredient in flour, are a source of sulfate (SO 2 4 ) free radicals under neutral pH. [4,5] Due to the higher oxidizing power of the sulfate radical (E 0 5 12.60 V) and longer half-life period over the OH radical, it has drawn attention in recent times as a powerful oxidizing agent. [6][7][8] Many chemical and cosmetic industries require persulfates as their key components [9] and workers often suffer from skin and respiratory diseases due to hypersensitive reactions. [10] Generally, the lower thermal energy at room temperature (25-508C) is not sufficient enough to activate persulfate, but generation of SO 2 4 radical is facilitated in presence of metal cations. Since proteins tend to lose their structure and function at high temperatures, [11] we have carried out the oxidation of ribonuclease A (RNase A) in presence of persulfate using cobalt (II) ion at 378C. [12] Cobalt is an integral component of vitamin B12 and thus physiologically found in most tissues mainly in liver and kidney. [13,14] Gill et al., reported the formation of oligomeric species (dimer, trimer, tetramer) when RNase A was exposed to KHSO 5 in presence of metal ions such as Ni 21 and Cu 21 and the high molecular weight species were identified using SDS-PAGE. [12] An important biomarker of oxidative stress is the dityrosine moiety generated due to the covalent linkage of two Tyr residues. [15] Dityrosine bond formation is an indication of aging, and disease related problems since it generally occurs during amyloid fibril formation, [16] Parkinson disease, [17] cerebrospinal fluid damage, [18] and also in the cataract of the eye lens. [19,20] D...

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Strategy for identification and detection of multiple oxidative modifications within proteins applied on persulfate‐oxidized hemoglobin and human serum albumin

Cited by 7 publications

References 30 publications

Targeted Proteomic Analyses of Nasal Lavage Fluid in Persulfate-Challenged Hairdressers with Bleaching Powder-Associated Rhinitis

Targeted Proteomic Analyses of Nasal Lavage Fluid in Persulfate-Challenged Hairdressers with Bleaching Powder-Associated Rhinitis

Efficiency of Trypsin Digestion for Mass-Spectrometry-Based Identification and Quantification of Oxidized Proteins: Evaluation of the Digestion of Oxidized Bovine Serum Albumin

β‐cyclodextrin encapsulated polyphenols as effective antioxidants

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