Site-specific nitrosoproteomic identification of endogenously S-nitrosylated proteins in Arabidopsis

Plant Physiol. 2015 Apr;167(4):1731-46. doi: 10.1104/pp.15.00026. Epub 2015 Feb 19.

Abstract

Nitric oxide (NO) regulates multiple developmental events and stress responses in plants. A major biologically active species of NO is S-nitrosoglutathione (GSNO), which is irreversibly degraded by GSNO reductase (GSNOR). The major physiological effect of NO is protein S-nitrosylation, a redox-based posttranslational modification mechanism by covalently linking an NO molecule to a cysteine thiol. However, little is known about the mechanisms of S-nitrosylation-regulated signaling, partly due to limited S-nitrosylated proteins being identified. In this study, we identified 1,195 endogenously S-nitrosylated peptides in 926 proteins from the Arabidopsis (Arabidopsis thaliana) by a site-specific nitrosoproteomic approach, which, to date, is the largest data set of S-nitrosylated proteins among all organisms. Consensus sequence analysis of these peptides identified several motifs that contain acidic, but not basic, amino acid residues flanking the S-nitrosylated cysteine residues. These S-nitrosylated proteins are involved in a wide range of biological processes and are significantly enriched in chlorophyll metabolism, photosynthesis, carbohydrate metabolism, and stress responses. Consistently, the gsnor1-3 mutant shows the decreased chlorophyll content and altered photosynthetic properties, suggesting that S-nitrosylation is an important regulatory mechanism in these processes. These results have provided valuable resources and new clues to the studies on S-nitrosylation-regulated signaling in plants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Arabidopsis / genetics
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / isolation & purification
  • Arabidopsis Proteins / metabolism
  • Cysteine / metabolism
  • Glutathione Reductase / genetics*
  • Glutathione Reductase / metabolism
  • Molecular Sequence Data
  • Nitric Oxide / metabolism*
  • Oxidation-Reduction
  • Protein Processing, Post-Translational*
  • Proteomics*
  • S-Nitrosoglutathione / metabolism*
  • Seedlings / genetics
  • Seedlings / metabolism
  • Sequence Alignment
  • Signal Transduction
  • Sulfhydryl Compounds / metabolism

Substances

  • Arabidopsis Proteins
  • Sulfhydryl Compounds
  • Nitric Oxide
  • S-Nitrosoglutathione
  • Glutathione Reductase
  • S-nitrosoglutathione reductase, Arabidopsis
  • Cysteine