Title: Protein microarray characterization of the S-nitrosoproteome.
Authors: 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 L; Dawson, Ted M
Published In Mol Cell Proteomics, (2014 Jan)
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 S-nitrosylated 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.
PubMed ID: 24105792
MeSH Terms: Humans; Nitric Oxide/metabolism*; Protein Array Analysis*; Protein Processing, Post-Translational/genetics*; Proteins/metabolism; Proteome*