A High-throughput Screening Compatible Assay for Activators and Inhibitors of Methionine Sulfoxide Reductase A

Overview

Journal Assay Drug Dev Technol

Specialties Chemistry
Pharmacology

Date 2010 Jun 3

PMID 20515413

Citations 4

Authors

David Brunell

Herbert Weissbach

Peter Hodder

Nathan Brot

Affiliations

Soon will be listed here.

Abstract

The methionine sulfoxide reductase (Msr) system has been shown to play an important role in protecting cells against oxidative damage. This family of enzymes can repair damage to proteins resulting from the oxidation of methionine residues to methionine sulfoxide, caused by reactive oxygen species. Previous genetic studies in animals have shown that increased levels of methionine sulfoxide reductase enzyme A (MsrA), an important member of the Msr family, can protect cells against oxidative damage and increase life span. A high-throughput screening (HTS) compatible assay has been developed to search for both activators and inhibitors of MsrA. The assay involves a coupled reaction in which the oxidation of NADPH is measured by either spectrophotometric or fluorometric analysis. Previous studies had shown that MsrA has a broad substrate specificity and can reduce a variety of methyl sulfoxide compounds, including dimethylsulfoxide (DMSO). Since the chemicals in the screening library are dissolved in DMSO, which would compete with any of the standard substrates used for the determination of MsrA activity, an assay has been developed that uses the DMSO that is the solvent for the compounds in the library as the substrate for the MsrA enzyme. A specific activator of MsrA could have important therapeutic value for diseases that involve oxidative damage, especially age-related diseases, whereas a specific inhibitor of MsrA would have value for a variety of research studies.

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References

Simeonov A, Jadhav A, Thomas C, Wang Y, Huang R, Southall N . Fluorescence spectroscopic profiling of compound libraries. J Med Chem. 2008; 51(8):2363-71. DOI: 10.1021/jm701301m. View

Moskovitz J, Weissbach H, Brot N . Cloning the expression of a mammalian gene involved in the reduction of methionine sulfoxide residues in proteins. Proc Natl Acad Sci U S A. 1996; 93(5):2095-9. PMC: 39915. DOI: 10.1073/pnas.93.5.2095. View

Simeonov A, Jadhav A, A Sayed A, Wang Y, Nelson M, Thomas C . Quantitative high-throughput screen identifies inhibitors of the Schistosoma mansoni redox cascade. PLoS Negl Trop Dis. 2008; 2(1):e127. PMC: 2217675. DOI: 10.1371/journal.pntd.0000127. View

Salmon A, Perez V, Bokov A, Jernigan A, Kim G, Zhao H . Lack of methionine sulfoxide reductase A in mice increases sensitivity to oxidative stress but does not diminish life span. FASEB J. 2009; 23(10):3601-8. PMC: 2747676. DOI: 10.1096/fj.08-127415. View

Levine R, Mosoni L, Berlett B, Stadtman E . Methionine residues as endogenous antioxidants in proteins. Proc Natl Acad Sci U S A. 1996; 93(26):15036-40. PMC: 26351. DOI: 10.1073/pnas.93.26.15036. View

Ejiri S, Weissbach H, Brot N . Reduction of methionine sulfoxide to methionine by Escherichia coli. J Bacteriol. 1979; 139(1):161-4. PMC: 216841. DOI: 10.1128/jb.139.1.161-164.1979. View

Marchetti M, Lee W, Cowell T, Wells T, Weissbach H, Kantorow M . Silencing of the methionine sulfoxide reductase A gene results in loss of mitochondrial membrane potential and increased ROS production in human lens cells. Exp Eye Res. 2006; 83(5):1281-6. PMC: 2831415. DOI: 10.1016/j.exer.2006.07.005. View

Minetti G, Balduini C, Brovelli A . Reduction of DABS-L-methionine-dl-sulfoxide by protein methionine sulfoxide reductase from polymorphonuclear leukocytes: stereospecificity towards the l-sulfoxide. Ital J Biochem. 1994; 43(6):273-83. View

Moskovitz J, Flescher E, Berlett B, Azare J, Poston J, Stadtman E . Overexpression of peptide-methionine sulfoxide reductase in Saccharomyces cerevisiae and human T cells provides them with high resistance to oxidative stress. Proc Natl Acad Sci U S A. 1998; 95(24):14071-5. PMC: 24328. DOI: 10.1073/pnas.95.24.14071. View

10.

Abrams W, WEINBAUM G, Weissbach L, Weissbach H, Brot N . Enzymatic reduction of oxidized alpha-1-proteinase inhibitor restores biological activity. Proc Natl Acad Sci U S A. 1981; 78(12):7483-6. PMC: 349292. DOI: 10.1073/pnas.78.12.7483. View

11.

Sagher D, Brunell D, Hejtmancik J, Kantorow M, Brot N, Weissbach H . Thionein can serve as a reducing agent for the methionine sulfoxide reductases. Proc Natl Acad Sci U S A. 2006; 103(23):8656-61. PMC: 1592241. DOI: 10.1073/pnas.0602826103. View

12.

Cochrane C, Spragg R, Revak S . Pathogenesis of the adult respiratory distress syndrome. Evidence of oxidant activity in bronchoalveolar lavage fluid. J Clin Invest. 1983; 71(3):754-61. PMC: 436926. DOI: 10.1172/jci110823. View

13.

Minond D, Saldanha S, Subramaniam P, Spaargaren M, Spicer T, Fotsing J . Inhibitors of VIM-2 by screening pharmacologically active and click-chemistry compound libraries. Bioorg Med Chem. 2009; 17(14):5027-37. PMC: 2759276. DOI: 10.1016/j.bmc.2009.05.070. View

14.

Brot N, Weissbach L, Werth J, Weissbach H . Enzymatic reduction of protein-bound methionine sulfoxide. Proc Natl Acad Sci U S A. 1981; 78(4):2155-8. PMC: 319302. DOI: 10.1073/pnas.78.4.2155. View

15.

Inglese J, Johnson R, Simeonov A, Xia M, Zheng W, Austin C . High-throughput screening assays for the identification of chemical probes. Nat Chem Biol. 2007; 3(8):466-79. DOI: 10.1038/nchembio.2007.17. View

16.

Shao B, Oda M, Bergt C, Fu X, Green P, Brot N . Myeloperoxidase impairs ABCA1-dependent cholesterol efflux through methionine oxidation and site-specific tyrosine chlorination of apolipoprotein A-I. J Biol Chem. 2006; 281(14):9001-4. DOI: 10.1074/jbc.C600011200. View

17.

Sagher D, Brunell D, Brot N, Vallee B, Weissbach H . Selenocompounds can serve as oxidoreductants with the methionine sulfoxide reductase enzymes. J Biol Chem. 2006; 281(42):31184-7. DOI: 10.1074/jbc.M606962200. View

18.

Hassouni M, Chambost J, Expert D, Van Gijsegem F, Barras F . The minimal gene set member msrA, encoding peptide methionine sulfoxide reductase, is a virulence determinant of the plant pathogen Erwinia chrysanthemi. Proc Natl Acad Sci U S A. 1999; 96(3):887-92. PMC: 15320. DOI: 10.1073/pnas.96.3.887. View

19.

Koc A, Gasch A, Rutherford J, Kim H, Gladyshev V . Methionine sulfoxide reductase regulation of yeast lifespan reveals reactive oxygen species-dependent and -independent components of aging. Proc Natl Acad Sci U S A. 2004; 101(21):7999-8004. PMC: 419546. DOI: 10.1073/pnas.0307929101. View

20.

Moskovitz J, Rahman M, Strassman J, Yancey S, Kushner S, Brot N . Escherichia coli peptide methionine sulfoxide reductase gene: regulation of expression and role in protecting against oxidative damage. J Bacteriol. 1995; 177(3):502-7. PMC: 176620. DOI: 10.1128/jb.177.3.502-507.1995. View