S‐Nitrosoglutathione in Rat Cerebellum: Identification and Quantification by Liquid Chromatography‐Mass Spectrometry
S‐Nitrosoglutathione in Rat Cerebellum: Identification and Quantification by Liquid Chromatography‐Mass Spectrometry
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DOI：
10.1046/j.1471-4159.1997.69062599.x
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发表时间：
1997-12
影响因子：
4.7
通讯作者：
I. Kluge;U. Gutteck-Amsler;M. Zollinger;K. Do
中科院分区：
文献类型：
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作者：
I. Kluge;U. Gutteck-Amsler;M. Zollinger;K. Do
摘要：鉴于信使一氧化氮(NO)的极不稳定和易被许多活性生化物种灭活，有人认为一些中间体化合物，如S亚硝硫醇，可能在稳定NO的同时保持其生物活性。为了验证这一假设，我们调查了大鼠大脑中是否存在谷胱甘肽的S-亚硝硫醇，谷胱甘肽是中枢神经系统中主要的低分子硫醇。[35S]半胱氨酸标记脑片小脑提取液的高效液相分析表明，大鼠脑内确实存在S-亚硝基谷胱甘肽(GSNO)。为了检测内源性GSNO，建立了一种基于液-质联用的检测方法。除了明确鉴定内源性GSNO外，该方法还允许以15N标记的GSNO([15N]-GSNO)为内标进行精确定量。成人小脑GSNO含量为15.4±1.4pmol/mg蛋白质。这是首次直接证明内源性GSNO存在于中枢神经系统。以GSNO的形式包装NO可能有助于其运输，延长其寿命，并将其靶向特定的效应器。
Abstract: Given the extreme lability and the facile inactivation of the messenger nitric oxide (NO) by many reactive biochemical species, it has been suggested that some intermediate compounds, for example, S‐nitrosothiols, may act to stabilize NO and at the same time to preserve its biological activity. To test this hypothesis, we investigated if the S‐nitrosothiol of glutathione, which is the predominant low molecular weight thiol in CNS, is present in the rat brain. The HPLC analysis of cerebellar extract from [35S]cysteine‐prelabeled slices suggested that S‐nitrosoglutathione (GSNO) was indeed present in rat brain. To detect endogenous GSNO, a methodology based on liquid chromatography‐mass spectrometry was developed. Besides an unequivocal identification of the endogenous GSNO, this method also permitted its precise quantification using 15N‐labeled GSNO ([15N]‐GSNO) as internal standard. GSNO level in adult cerebellum amounts to 15.4 ± 1.4 pmol/mg of protein. This is the first direct demonstration of the presence of endogenous GSNO in CNS. The packaging of NO in the form of GSNO might serve to facilitate its transport, prolong its life, and target its delivery to specific effectors.