Strategies for profiling native S-nitrosylation.

Strategies for profiling native S-nitrosylation.
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DOI:
10.1002/bip.22342
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发表时间:
2014-02
期刊:
影响因子:
2.9
通讯作者:
Martin, Brent R.
Martin, Brent R.
中科院分区:
生物学4区
文献类型:
--
作者:
Majmudar, Jaimeen D.;Martin, Brent R.

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半胱氨酸是一种独特的反应性氨基酸,能够进行亲核和氧化翻译后修饰。一种这样的氧化反应涉及通过气态第二信使一氧化氮(NO)共价修饰半胱氨酸,称为S-亚硝基化(SNO)。这种动态的翻译后修饰参与了所有致癌界蛋白质的氧化还原调节。在哺乳动物中,一氧化氮合酶的钙依赖性激活触发一氧化氮的局部释放,其激活附近的鸟苷酸环化酶和cGMP依赖性途径。同时,可扩散的一氧化氮可以局部修饰氧化还原活性细胞硫醇,在功能上调节许多氧化还原敏感酶。异常S-亚硝基化与许多疾病的病理学有关,包括神经变性、炎症和中风。在这篇综述中,我们讨论了目前的方法来标记位点的S-亚硝基化的生化分析。最流行的方法包括一系列的生化步骤,以掩盖游离硫醇,然后选择性亚硝基硫醇还原和捕获。其他新出现的方法包括基于机理的膦探针和汞富集化学。通过将新的富集方法与高分辨率质谱法相结合,蛋白质亚硝基化的大规模分析突出了氧化调节的新途径。
Cysteine is a uniquely reactive amino acid, capable of undergoing both nucleophlilic and oxidative post-translational modifications. One such oxidation reaction involves the covalent modification of cysteine via the gaseous second messenger nitric oxide (NO), termed S-nitrosylation (SNO). This dynamic post-translational modification is involved in the redox regulation of proteins across all phylogenic kingdoms. In mammals, calcium-dependent activation of nitric oxide synthase triggers the local release of nitric oxide, which activates nearby guanylyl cyclases and cGMP-dependent pathways. In parallel, diffusible nitric oxide can locally modify redox active cellular thiols, functionally modulating many redox sensitive enzymes. Aberrant S-nitrosylation is implicated in the pathology of many diseases, including neurodegeneration, inflammation, and stroke. In this review, we discuss current methods to label sites of S-nitrosylation for biochemical analysis. The most popular method involves a series of biochemical steps to mask free thiols followed by selective nitrosothiol reduction and capture. Other emerging methods include mechanism-based phosphine probes and mercury enrichment chemistry. By bridging new enrichment approaches with high-resolution mass spectrometry, large-scale analysis of protein nitrosylation has highlighted new pathways of oxidative regulation.
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