Protein microarray characterization of the S-nitrosoproteome

Yun Il Lee, Daniel Giovinazzo, Ho Chul Kang, Yunjong Lee, Jun Seop Jeong, Paschalis Thomas Doulias, Zhi Xie, Jianfei Hu, Mehdi Ghasemi, Harry Ischiropoulos, Jiang Qian, Heng Zhu, Seth Blackshaw, Valina Dawson, Ted M Dawson

Research output: Contribution to journalArticle

Abstract

Nitric oxide (NO) mediates a substantial part of its physiologic functions via S-nitrosylation, however the cellular substrates for NO-mediated S-nitrosylation are largely unknown. Here we describe the S-nitrosoproteome using a high-density protein microarray chip containing 16,368 unique human proteins. We identified 834 potentially Snitrosylated human proteins. Using a unique and highly specific labeling and affinity capture of S-nitrosylated proteins, 138 cysteine residues on 131 peptides in 95 proteins were determined, defining critical sites of NO's actions. Of these cysteine residues 113 are novel sites of S-nitrosylation. A consensus sequence motif from these 834 proteins for S-nitrosylation was identified, suggesting that the residues flanking the S-nitrosylated cysteine are likely to be the critical determinant of whether the cysteine is S-nitrosylated. We identify eight ubiquitin E3 ligases, RNF10, RNF11, RNF41, RNF141, RNF181, RNF208, WWP2, and UBE3A, whose activities are modulated by S-nitrosylation, providing a unique regulatory mechanism of the ubiquitin proteasome system. These results define a new and extensive set of proteins that are susceptible to NO regulation via S-nitrosylation. Similar approaches could be used to identify other post-translational modification proteomes. Molecular & Cellular Proteomics 13: 10.1074/ mcp.M113.032235, 63-72, 2014.

Original languageEnglish (US)
Pages (from-to)63-72
Number of pages10
JournalMolecular and Cellular Proteomics
Volume13
Issue number1
DOIs
StatePublished - Jan 2014

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Protein Array Analysis
Microarrays
Cysteine
Nitric Oxide
Protein S
Proteins
Ubiquitin-Protein Ligases
Consensus Sequence
Proteasome Endopeptidase Complex
Proteome
Post Translational Protein Processing
Ubiquitin
Proteomics
Labeling
Peptides
Substrates

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Cite this

Lee, Y. I., Giovinazzo, D., Kang, H. C., Lee, Y., Jeong, J. S., Doulias, P. T., ... Dawson, T. M. (2014). Protein microarray characterization of the S-nitrosoproteome. Molecular and Cellular Proteomics, 13(1), 63-72. https://doi.org/10.1074/mcp.M113.030254

Protein microarray characterization of the S-nitrosoproteome. / Lee, Yun Il; Giovinazzo, Daniel; Kang, Ho Chul; Lee, Yunjong; Jeong, Jun Seop; Doulias, Paschalis Thomas; Xie, Zhi; Hu, Jianfei; Ghasemi, Mehdi; Ischiropoulos, Harry; Qian, Jiang; Zhu, Heng; Blackshaw, Seth; Dawson, Valina; Dawson, Ted M.

In: Molecular and Cellular Proteomics, Vol. 13, No. 1, 01.2014, p. 63-72.

Research output: Contribution to journalArticle

Lee, YI, Giovinazzo, D, Kang, HC, Lee, Y, Jeong, JS, Doulias, PT, Xie, Z, Hu, J, Ghasemi, M, Ischiropoulos, H, Qian, J, Zhu, H, Blackshaw, S, Dawson, V & Dawson, TM 2014, 'Protein microarray characterization of the S-nitrosoproteome', Molecular and Cellular Proteomics, vol. 13, no. 1, pp. 63-72. https://doi.org/10.1074/mcp.M113.030254
Lee YI, Giovinazzo D, Kang HC, Lee Y, Jeong JS, Doulias PT et al. Protein microarray characterization of the S-nitrosoproteome. Molecular and Cellular Proteomics. 2014 Jan;13(1):63-72. https://doi.org/10.1074/mcp.M113.030254
Lee, Yun Il ; Giovinazzo, Daniel ; Kang, Ho Chul ; Lee, Yunjong ; Jeong, Jun Seop ; Doulias, Paschalis Thomas ; Xie, Zhi ; Hu, Jianfei ; Ghasemi, Mehdi ; Ischiropoulos, Harry ; Qian, Jiang ; Zhu, Heng ; Blackshaw, Seth ; Dawson, Valina ; Dawson, Ted M. / Protein microarray characterization of the S-nitrosoproteome. In: Molecular and Cellular Proteomics. 2014 ; Vol. 13, No. 1. pp. 63-72.
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