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Identification of S-nitrosylated targets of thioredoxin using a quantitative proteomic approach.

Publication ,  Journal Article
Benhar, M; Thompson, JW; Moseley, MA; Stamler, JS
Published in: Biochemistry
August 17, 2010

Reversible protein cysteine nitrosylation (S-nitrosylation) is a common mechanism utilized in signal transduction and other diverse cellular processes. Protein denitrosylation is largely mediated by cysteine denitrosylases, but the functional scope and significance of these enzymes are incompletely defined, in part due to limited information on their cognate substrates. Here, using Jurkat cells, we employed stable isotope labeling by amino acids in cell culture (SILAC), coupled to the biotin switch technique and mass spectrometry, to identify 46 new substrates of one denitrosylase, thioredoxin 1. These substrates are involved in a wide range of cellular functions including cytoskeletal organization, cellular metabolism, signal transduction, and redox homeostasis. We also identified multiple S-nitrosylated proteins that are not substrates of thioredoxin 1. A verification of our principal findings was made in a second cell type (RAW264.7 cells). Our results point to thioredoxin 1 as a major protein denitrosylase in mammalian cells and demonstrate the utility of quantitative proteomics for large-scale identification of denitrosylase substrates.

Duke Scholars

Published In

Biochemistry

DOI

EISSN

1520-4995

Publication Date

August 17, 2010

Volume

49

Issue

32

Start / End Page

6963 / 6969

Location

United States

Related Subject Headings

  • Thioredoxins
  • Tandem Mass Spectrometry
  • S-Nitrosothiols
  • Proteomics
  • Models, Theoretical
  • Mice
  • Jurkat Cells
  • Isotope Labeling
  • Humans
  • Chromatography, Liquid
 

Citation

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Benhar, M., Thompson, J. W., Moseley, M. A., & Stamler, J. S. (2010). Identification of S-nitrosylated targets of thioredoxin using a quantitative proteomic approach. Biochemistry, 49(32), 6963–6969. https://doi.org/10.1021/bi100619k
Benhar, Moran, J Will Thompson, M Arthur Moseley, and Jonathan S. Stamler. “Identification of S-nitrosylated targets of thioredoxin using a quantitative proteomic approach.Biochemistry 49, no. 32 (August 17, 2010): 6963–69. https://doi.org/10.1021/bi100619k.
Benhar M, Thompson JW, Moseley MA, Stamler JS. Identification of S-nitrosylated targets of thioredoxin using a quantitative proteomic approach. Biochemistry. 2010 Aug 17;49(32):6963–9.
Benhar, Moran, et al. “Identification of S-nitrosylated targets of thioredoxin using a quantitative proteomic approach.Biochemistry, vol. 49, no. 32, Aug. 2010, pp. 6963–69. Pubmed, doi:10.1021/bi100619k.
Benhar M, Thompson JW, Moseley MA, Stamler JS. Identification of S-nitrosylated targets of thioredoxin using a quantitative proteomic approach. Biochemistry. 2010 Aug 17;49(32):6963–6969.
Journal cover image

Published In

Biochemistry

DOI

EISSN

1520-4995

Publication Date

August 17, 2010

Volume

49

Issue

32

Start / End Page

6963 / 6969

Location

United States

Related Subject Headings

  • Thioredoxins
  • Tandem Mass Spectrometry
  • S-Nitrosothiols
  • Proteomics
  • Models, Theoretical
  • Mice
  • Jurkat Cells
  • Isotope Labeling
  • Humans
  • Chromatography, Liquid