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 journalArticlepeer-review


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.

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

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology


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