Protein S-nitrosylation and denitrosylation in the mouse spinal cord upon injury of the sciatic nerve

J Proteomics. 2012 Jul 16;75(13):3987-4004. doi: 10.1016/j.jprot.2012.05.006. Epub 2012 May 14.


Nitric oxide is a pain signaling molecule and exerts its influence through two primary pathways: by stimulation of soluble guanylylcyclase and by direct S-nitrosylation (SNO) of target proteins. We assessed in the spinal cord the SNO-proteome with two methods, two-dimensional S-nitrosothiol difference gel electrophoresis (2D SNO-DIGE) and SNO-site identification (SNOSID) at baseline and 24h after sciatic nerve injury with/without pretreatment with the nitric oxide synthase inhibitor L-NG-nitroarginine methyl ester (L-NAME). After nerve injury, SNO-DIGE revealed 30 proteins with increased and 23 proteins with decreased S-nitrosylation. SNO-sites were identified for 17 proteins. After sham surgery only 3 proteins were up-nitrosylated. L-NAME pretreatment substantially reduced both constitutive and nerve injury evoked up-S-nitrosylation. For the top candidates S-nitrosylation was confirmed with the biotin switch technique and time course analyses at 1 and 7days showed that SNO modifications of protein disulfide isomerase, glutathione synthase and peroxiredoxin-6 had returned to baseline within 7days whereas S-nitrosylation of mitochondrial aconitase 2 was further increased. The identified SNO modified proteins are involved in mitochondrial function, protein folding and transport, synaptic signaling and redox control. The data show that nitric oxide mediated S-nitrosylation contributes to the nerve injury-evoked pathology in nociceptive signaling pathways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aconitate Hydratase / metabolism
  • Animals
  • Glutathione Synthase / metabolism
  • Mice
  • NG-Nitroarginine Methyl Ester / pharmacology
  • Nitric Oxide Synthase / antagonists & inhibitors
  • Oxidation-Reduction
  • Peroxiredoxin VI / metabolism
  • Protein Disulfide-Isomerases / metabolism
  • Proteomics / methods
  • S-Nitrosothiols / metabolism*
  • Sciatic Nerve / injuries*
  • Signal Transduction / physiology
  • Spinal Cord / metabolism*
  • Two-Dimensional Difference Gel Electrophoresis


  • S-Nitrosothiols
  • Peroxiredoxin VI
  • Nitric Oxide Synthase
  • Aconitate Hydratase
  • aconitase 2, mouse
  • Protein Disulfide-Isomerases
  • Glutathione Synthase
  • NG-Nitroarginine Methyl Ester