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. 2019 Jan 14;9(1):115.
doi: 10.1038/s41598-018-35566-2.

Cysteine Disulfides (Cys-ss-X) as Sensitive Plasma Biomarkers of Oxidative Stress

Xiaoyun Fu  1   2 , Shelby A Cate  3 , Melissa Dominguez  3 , Warren Osborn  3 , Tahsin Özpolat  3 , Barbara A Konkle  3   4 , Junmei Chen  3 , José A López  5   6
Affiliations

Cysteine Disulfides (Cys-ss-X) as Sensitive Plasma Biomarkers of Oxidative Stress

Xiaoyun Fu et al. Sci Rep. .

Abstract

We developed a high-throughput mass spectrometry-based method to simultaneously quantify numerous small-molecule thiols and disulfides in blood plasma. Application of this assay to analyze plasma from patients with known oxidative stress (sickle cell disease and sepsis) and from a patient with sickle cell disease treated with the antioxidant N-acetylcysteine suggests that cysteine disulfides, in particular protein-bound cysteine, serve as sensitive plasma biomarkers for the extent of oxidative stress and effectiveness of antioxidant treatment.

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Conflict of interest statement

X.F., S.A.C., J.A.L., J.C., B.A.K. and BloodworksNW have filed a patent regarding biomarkers and methods presented in this paper (application number: WO2016015060A1, status: pending). Other authors declare no conflict of interests.

Figures

Figure 1
Figure 1
Quantification of glutathione and cysteine species by LC-MS/MS-MRM. (a) Workflow for sample processing for MS analysis. (b) A representative UPLC chromatogram for a panel of disulfides and NEM-alkylated thiols.
Figure 2
Figure 2
Cysteine, not glutathione, is the major thiol-containing compound in plasma. (a) Plasma concentrations of reduced and total forms of glutathione, cysteine (Cys), Cys-Gly, γ-Glu-Cys, and homocysteine (Hcy). (b) Concentrations of Cys either in reduced form or disulfide-bonded adducts measured in plasma. Data in panels (a) and (b) are from 11 healthy donors.
Figure 3
Figure 3
Protein-bound cysteine (protein-ss-Cys) in plasma is a biomarker for oxidative stress. (a,b) Oxidization of plasma by H2O2 and HOCl. The concentrations of protein-ss-Cys increased in plasma as oxidant concentration increased (a) while reduced Cys decreased (b). (c) Compared to healthy donors (n = 12), patients with SCD (n = 9), or sepsis (n = 6) had significantly elevated protein-ss-Cys and total Cys concentrations in plasma. (d) The concentration of protein-ss-Cys correlated positively with the total Cys concentration in plasma. For sepsis, the data are derived from 6 samples from 5 patients and the statistics are calculated based on n = 6. The 2 samples from one patient (patient 3 in Supplementary Table 3) were taken 3 days apart.
Figure 4
Figure 4
Changes of plasma Cys species in an SCD patient received NAC infusion. An SCD patient was infused with NAC, 150 mg/kg over 1 hour. (a) Concentrations of NAC and its disulfide-bonded forms measured at the end of the infusion. (b) Time course of concentrations of total, reduced, and protein-bound Cys before and after NAC infusion.
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References

    1. Dalle-Donne I, Rossi R, Colombo R, Giustarini D, Milzani A. Biomarkers of oxidative damage in human disease. Clin. Chem. 2006;52:601–623. doi: 10.1373/clinchem.2005.061408. - DOI - PubMed
    1. Buglak, N. E., Batrakova, E. V., Mota, R. & Bahnson, E. S. M. Insights on localized and systemic delivery of redox-based therapeutics. Oxid. Med. Cell Longev. 10.1155/2018/2468457 (2018). - PMC - PubMed
    1. Rossi R, Dalle-Donne I, Milzani A, Giustarini D. Oxidized forms of glutathione in peripheral blood as biomarkers of oxidative stress. Clin. Chem. 2006;52:1406–1414. doi: 10.1373/clinchem.2006.067793. - DOI - PubMed
    1. Rahman I, Kode A, Biswas SK. Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat. Protoc. 2006;1:3159–3165. doi: 10.1038/nprot.2006.378. - DOI - PubMed
    1. Kleinman WA, Richie JP., Jr. Status of glutathione and other thiols and disulfides in human plasma. Biochem. Pharmacol. 2000;60:19–29. doi: 10.1016/S0006-2952(00)00293-8. - DOI - PubMed

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