Mass spectrometry of fluorocarbon-labeled glycosphingolipids.
Mass spectrometry of fluorocarbon-labeled glycosphingolipids.
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DOI：
10.1002/jms.1734
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发表时间：
2010-05
期刊：
影响因子：
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通讯作者：
Yunsen Li;Emma Arigi;Heather Eichert;S. Levery
中科院分区：
文献类型：
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作者：
Yunsen Li;Emma Arigi;Heather Eichert;S. Levery
开发了一种用于产生对氟碳相具有高亲和力的鞘糖脂（GSL）的新型氟碳衍生物的方法，并研究了它们在基于质谱（MS）的鞘糖脂组学方法中的潜在应用。鞘脂神经酰胺N-脱酰基酶（SCDase）用于从神经酰胺部分去除脂肪酸，之后在类鞘氨醇的游离胺处掺入富含氟碳的取代基（F-Tag）。在最初的试验中，中性GSL，globotriaosylceramide（Gb（3）Cer），三个纯化的牛脑神经节苷脂，和四个真菌糖基肌醇磷酸神经酰胺（GIPC）的脱N-酰化，衍生的原型F-Tags，并通过固相萃取回收的氟碳衍生的二氧化硅（F-SPE）。本文首次证明了SCDase治疗GIPC的疗效。对于F-标记的Gb（3）Cer和纯化的神经节苷脂，建立了与随后的per-N，O-甲基化的相容性，并且获得了与所有预期产物一致的广泛质谱（MS（1）和MS（2））。然后在从牛脑中提取的粗神经节苷脂混合物上证明了F-标记的衍生物用于全面的基于MS的分析应用的潜在用途。最后，一个简单的微阵列形式的试验证明了F-标记的G（M1）神经节苷脂固定到氟玻璃表面，聚糖完整，可与霍乱毒素B链的荧光衍生物相互作用。因此，所描述的方法提供了一种新的途径，用于快速GSL回收或清除，可能与各种平台兼容的质谱分析和结构分析，以及功能相互作用的平行分析。
A method for generation of novel fluorocarbon derivatives of glycosphingolipids (GSLs) with high affinity for fluorocarbon phases has been developed, and their potential applications to mass spectrometry (MS)-based methodologies for glycosphingolipidomics have been investigated. Sphingolipid ceramide N-deacylase (SCDase) is used to remove the fatty acid from the ceramide moiety, after which a fluorocarbon-rich substituent (F-Tag) is incorporated at the free amine of the sphingoid. In initial trials, a neutral GSL, globotriaosylceramide (Gb(3)Cer), three purified bovine brain gangliosides, and four fungal glycosylinositol phosphorylceramides (GIPCs) were de-N-acylated, derivatized by prototype F-Tags, and recovered by solid phase extraction on fluorocarbon-derivatized silica (F-SPE). The efficacy of SCDase treatment of GIPCs was here demonstrated for the first time. Compatibility with subsequent per-N,O-methylation was established for the F-tagged Gb(3) Cer and purified gangliosides, and extensive mass spectra (MS(1) and MS(2)) consistent with all of the expected products were acquired. The potential use of F-tagged derivatives for a comprehensive MS based profiling application was then demonstrated on a crude ganglioside mixture extracted from bovine brain. Finally, a simple trial in microarray format demonstrated fixation of F-tagged G(M1) ganglioside to a fluorous glass surface, with the glycan intact and available for interaction with a fluorescent derivative of cholera toxin B chain. The methods described thus provide a new avenue for rapid GSL recovery or cleanup, potentially compatible with a variety of platforms for mass spectrometric profiling and structure analysis, as well as parallel analysis of functional interactions.