Fast screening of highly glycosylated plant sphingolipids by tandem mass spectrometry
Fast screening of highly glycosylated plant sphingolipids by tandem mass spectrometry
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DOI：
10.1002/rcm.5206
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发表时间：
2011-10-30
影响因子：
2
通讯作者：
Schmitter, Jean-Marie
中科院分区：
文献类型：
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作者：
Bure, Corinne;Cacas, Jean-Luc;Schmitter, Jean-Marie
糖基肌醇磷酸神经酰胺（GIPC）是植物组织中的主要鞘脂，其结构表征是了解其生理功能的关键一步。在优化其提取后，许多植物GIPC已通过质谱法表征。负离子的基质辅助激光解吸/电离质谱（MALDI-MS）全扫描分析提供了GIPC分布的快速概述。在两种植物模型中观察到明显的差异：在烟草BY-2细胞提取物中检测到6个含有2 - 7个糖单元的GIPC系列，而从A.拟南芥细胞培养物和叶片的多样性较小，主要是只含有两个糖单位的物种。在A. thaliana和120在烟草BY-2细胞中的表达。MALDI-MS/MS光谱提供了关于神经酰胺部分、极性头部以及糖单元的数量和类型的详细结构信息。一旦从GIPC释放，通过GC/MS分析脂肪酸链和长链碱基，以验证MALDI-MS/MS方法考虑了所有GIPC系列。ESI-MS/MS为同量异位物质和脂肪酸链的鉴定提供了补充信息。这种方法，主要依赖于MALDI-MS/MS，应该打开新的途径，以确定鞘糖脂和膜组织之间的结构-功能关系。版权所有（C）2011约翰威利父子有限公司
The structural characterization of Glycosyl-Inositol-Phospho-Ceramides (GIPCs), which are the main sphingolipids of plant tissues, is a critical step towards the understanding of their physiological function. After optimization of their extraction, numerous plant GIPCs have been characterized by mass spectrometry. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) full scan analysis of negative ions provides a quick overview of GIPC distribution. Clear differences were observed for the two plant models studied: six GIPC series bearing from two to seven saccharide units were detected in tobacco BY-2 cell extracts, whereas GIPCs extracted from A. thaliana cell cultures and leaves were less diverse, with a dominance of species containing only two saccharide units. The number of GIPC species was around 50 in A. thaliana and 120 in tobacco BY-2 cells. MALDI-MS/MS spectra gave access to detailed structural information relative to the ceramide moiety, the polar head, as well as the number and types of saccharide units. Once released from GIPCs, fatty acid chains and long-chain bases were analyzed by GC/MS to verify that all GIPC series were taken into account by the MALDI-MS/MS approach. ESI-MS/MS provided complementary information for the identification of isobaric species and fatty acid chains. Such a methodology, mostly relying on MALDI-MS/MS, should open new avenues to determine structure-function relationships between glycosphingolipids and membrane organization. Copyright (C) 2011 John Wiley & Sons, Ltd.