Protein S-nitrosylation and denitrosylation in the mouse spinal cord upon injury of the sciatic nerve |
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Authors: | Scheving Reynir Wittig Ilka Heide Heinrich Albuquerque Boris Steger Mirco Brandt Ulrich Tegeder Irmgard |
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Affiliation: | 1. Institut für Klinische Pharmakologie, Klinikum der Goethe-Universität Frankfurt, 60590 Frankfurt am Main, Germany;2. Molekulare Bioenergetik, Zentrum der Biologischen Chemie, Cluster of Excellence Frankfurt Macromolecular Complexes, Goethe-Universität Frankfurt, 60590 Frankfurt am Main, Germany |
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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. |
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Keywords: | 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 |
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