title: Protein S-nitrosylation and denitrosylation in the mouse spinal cord upon injury of the sciatic nerve.
abstract: 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.
authors.affiliation: Institut für Klinische Pharmakologie, Klinikum der Goethe-Universität Frankfurt, 60590 Frankfurt am Main, Germany.
authors.forename: Reynir
authors.initials: R
authors.lastname: Scheving
authors.forename: Ilka
authors.initials: I
authors.lastname: Wittig
authors.forename: Heinrich
authors.initials: H
authors.lastname: Heide
authors.forename: Boris
authors.initials: B
authors.lastname: Albuquerque
authors.forename: Mirco
authors.initials: M
authors.lastname: Steger
authors.forename: Ulrich
authors.initials: U
authors.lastname: Brandt
authors.forename: Irmgard
authors.initials: I
authors.lastname: Tegeder
mesh_terms: D000154:Aconitate Hydratase / Q000378:metabolism; D000818:Animals; D005981:Glutathione Synthase / Q000378:metabolism; D051379:Mice; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D010084:Oxidation-Reduction; D054465:Peroxiredoxin VI / Q000378:metabolism; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D026403:S-Nitrosothiols / Q000378:metabolism*; D012584:Sciatic Nerve / Q000293:injuries*; D015398:Signal Transduction / Q000502:physiology; D013116:Spinal Cord / Q000378:metabolism*; D057825:Two-Dimensional Difference Gel Electrophoresis
chemical_list: D026403:S-Nitrosothiols; D054465:Peroxiredoxin VI; D019001:Nitric Oxide Synthase; D000154:Aconitate Hydratase; C543483:aconitase 2, mouse; D019704:Protein Disulfide-Isomerases; D005981:Glutathione Synthase; D019331:NG-Nitroarginine Methyl Ester