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Nitric Oxide pp 253-266 | Cite as

Biotin Switch Processing and Mass Spectrometry Analysis of S-Nitrosated Thioredoxin and Its Transnitrosation Targets

  • Changgong Wu
  • Tong Liu
  • Yan Wang
  • Lin Yan
  • Chuanlong Cui
  • Annie Beuve
  • Hong LiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1747)

Abstract

S-Nitrosation is a key posttranslational modification in regulating proteins in both normal physiology and diverse human diseases. To identify novel therapies for human diseases linked to oxidative and nitrosative stress, understanding how cells control S-nitrosation specificity could be critical. Among the enzymes known to control S-nitrosation of proteins, thioredoxin 1 (Trx1), a conserved disulfide reductase, transnitrosates and denitrosates distinct sets of target proteins. To recognize the function of Trx1 in both normal and dysfunctional cells, S-nitrosation targets of Trx1 in different cells need to be identified. However, S-nitrosation is usually too labile to be detected directly by mass spectrometry (MS). Here we present two optimized MS techniques to identify S-nitrosated Trx1 and its transnitrosation targets, using both direct and indirect MS methods.

Key words

S-nitrosation Thioredoxin Transnitrosation Mass spectrometry 

Notes

Acknowledgments

This chapter is produced with a grant support from the NIH-National Institute of General Medical Sciences (R01GM112415, to A.B. and H.L.), and the Orbitrap MS described here was purchased with a grant from the NIH-National Institute of Neurological Disorders and Stroke (P30NS046593).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Changgong Wu
    • 1
  • Tong Liu
    • 1
  • Yan Wang
    • 2
  • Lin Yan
    • 1
  • Chuanlong Cui
    • 1
  • Annie Beuve
    • 3
  • Hong Li
    • 1
    Email author
  1. 1.Center for Advanced Proteomics Research and Department of Microbiology, Biochemistry and Molecular GeneticsRutgers, New Jersey Medical SchoolNewarkUSA
  2. 2.School of PharmacyShanghai Jiao Tong UniversityShanghaiP. R. China
  3. 3.Department of Pharmacology, Physiology and NeuroscienceRutgers, New Jersey Medical SchoolNewarkUSA

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