Involvement of S-nitrosylation of actin in inhibition of neurotransmitter release by nitric oxide
Involvement of S-nitrosylation of actin in inhibition of neurotransmitter release by nitric oxide
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DOI：
10.1186/1744-8069-5-58
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发表时间：
2009-09-29
期刊：
影响因子：
3.3
通讯作者：
Ito, Seiji
中科院分区：
文献类型：
--
作者：
Lu, Jingshan;Katano, Tayo;Ito, Seiji
背景资料：可扩散信使一氧化氮（NO）在痛觉传递中的作用仍有争议，是促伤害性还是抗伤害性的。S-亚硝基化是蛋白质中选择性半胱氨酸残基的可逆翻译后修饰，已成为NO作为信号分子的重要机制。脊髓中S-亚硝基化的发生及其可能调节疼痛传递的靶点仍不清楚。“生物素开关”法和基质辅助激光解吸电离飞行时间质谱法被用于鉴定S-亚硝基化的蛋白质。结果：在这里，我们表明，肌动蛋白是一个主要的蛋白质S-亚硝基化的脊髓由NO供体，S-亚硝基-N-乙酰基-DL-青霉胺（SNAP）。有趣的是，肌动蛋白的S-亚硝基化，更多的S2馏分比P2馏分的脊髓匀浆。用SNAP处理PC 12细胞引起肌动蛋白的快速S-亚硝基化，并抑制细胞中多巴胺的释放。就像细胞松弛素B，解聚肌动蛋白，SNAP减少了膜下的丝状肌动蛋白细胞骨架的数量。可溶性鸟苷酸环化酶和cGMP依赖性蛋白激酶抑制剂对多巴胺释放的抑制没有减弱。结论：本研究表明，肌动蛋白是一种主要的S-亚硝基化蛋白在脊髓，并表明，NO直接调节神经递质释放的S-亚硝基化除了众所周知的磷酸化cGMP依赖性蛋白激酶。
Background: The role of the diffusible messenger nitric oxide (NO) in the regulation of pain transmission is still a debate of matter, pro-nociceptive and/or anti-nociceptive. S-Nitrosylation, the reversible post-translational modification of selective cysteine residues in proteins, has emerged as an important mechanism by which NO acts as a signaling molecule. The occurrence of S-nitrosylation in the spinal cord and its targets that may modulate pain transmission remain unclarified. The "biotin-switch" method and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were employed for identifying S-nitrosylated proteins.Results: Here we show that actin was a major protein S-nitrosylated in the spinal cord by the NO donor, S-nitroso-N-acetyl-DL-penicillamine (SNAP). Interestingly, actin was S-nitrosylated, more in the S2 fraction than in the P2 fraction of the spinal homogenate. Treatment of PC12 cells with SNAP caused rapid S-nitrosylation of actin and inhibited dopamine release from the cells. Just like cytochalasin B, which depolymerizes actin, SNAP decreased the amount of filamentous actin cytoskeleton just beneath the membrane. The inhibition of dopamine release was not attenuated by inhibitors of soluble guanylyl cyclase and cGMP-dependent protein kinase.Conclusion: The present study demonstrates that actin is a major S-nitrosylated protein in the spinal cord and suggests that NO directly regulates neurotransmitter release by S-nitrosylation in addition to the well-known phosphorylation by cGMP-dependent protein kinase.