Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice
<p>Abstract</p> <p>Background</p> <p><it>S</it>-Nitrosylation, the reversible post-translational modification of reactive cysteine residues in proteins, has emerged as an important mechanism by which NO acts as a signaling molecule. We recently demonstrated...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Book |
| Published: |
SAGE Publishing,
2011-12-01T00:00:00Z.
|
| Subjects: | |
| Online Access: | Connect to this object online. |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Summary: | <p>Abstract</p> <p>Background</p> <p><it>S</it>-Nitrosylation, the reversible post-translational modification of reactive cysteine residues in proteins, has emerged as an important mechanism by which NO acts as a signaling molecule. We recently demonstrated that actin is a major <it>S</it>-nitrosylated protein in the spinal cord and suggested that NO directly attenuates dopamine release from PC12 cells by causing the breakdown of F-actin. However, the occurrence of <it>S</it>-nitrosylation of actin remained unclarified in animal pain model. Kinetic analysis of <it>S</it>-nitrosylation of actin in the present study was made by using NO-generating donors. The biotin-switch assay and purification on streptavidin-agarose were employed for identification of <it>S</it>-nitrosylated actin.</p> <p>Results</p> <p>Dopamine release from PC12 cells was markedly attenuated by NOR1 (<it>t</it><sub>1/2 </sub>= 1.8 min) and much less by NOR3 (<it>t</it><sub>1/2 </sub>= 30 min), but not by <it>S</it>-nitroso-glutathione, an endogenous NO donor. A membrane-permeable cGMP analogue could not substitute for NOR1 as a suppressor nor could inhibitors of soluble guanylate cyclase and cGMP-dependent protein kinase attenuate the suppression. <it>S</it>-Nitrosylated actin was detected by the biotin-switch assay at 5 min after the addition of NOR1. Consistent with the kinetic analysis, actin in the spinal cord was rapidly and maximally <it>S</it>-nitrosylated in an inflammatory pain model at 5 min after the injection of 2% formalin into the hind paws. <it>In vivo </it>patch-clamp recordings of the spinal dorsal horn, NOR3 showed an inhibitory action on inhibitory synaptic transmission in interneurons of the substantia gelatinosa.</p> <p>Conclusions</p> <p>The present study demonstrates that rapid <it>S</it>-nitrosylation of actin occurred <it>in vitro </it>in the presence of exogenous NO-generating donors and <it>in vivo </it>in inflammatory pain model mice. Our data suggest that, in addition to the well-known cGMP-dependent protein kinase pathway, <it>S</it>-nitrosylation is involved in pain transmission via disinhibition of inhibitory neurons.</p> |
|---|---|
| Item Description: | 10.1186/1744-8069-7-101 1744-8069 |