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

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• 作者：Reynir Scheving ; Ilka Wittig ; Heinrich Heide ; Boris Albuquerque ; Mirco Steger ; Ulrich Br ; t ; Irmgard Tegeder
• 关键词：NO ; nitric oxide ; SNO ; S-nitrosylation or thiol-nitrosylation ; SNI ; spared nerve injury ; 2D SNO-DIGE ; S-nitrosylation two-dimensional gel electrophoresis ; SNOSID ; SNO site identification ; BST ; biotin switch technique
• 刊名：Journal of Proteomics
• 出版年：2012
• 出版时间：16 July, 2012
• 年：2012
• 卷：75
• 期：13
• 页码：3987-4004
• 全文大小：970 K

文摘

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 24 h 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 7 days showed that SNO modifications of protein disulfide isomerase, glutathione synthase and peroxiredoxin-6 had returned to baseline within 7 days 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.