Sphingolipidomics: methods for the comprehensive analysis of sphingolipids.
Sphingolipidomics: methods for the comprehensive analysis of sphingolipids.
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DOI：
10.1016/j.jchromb.2008.12.057
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发表时间：
2009-09-15
期刊：
影响因子：
--
通讯作者：
Sullards MC
中科院分区：
文献类型：
--
作者：
Haynes CA;Allegood JC;Park H;Sullards MC
鞘脂类包含高度多样和复杂的分子类别，它们既是细胞膜的结构成分，又是能够在哺乳动物细胞中引发细胞凋亡、分化、趋化性和其他反应的信号分子。为了阐明鞘脂类在特定生物环境中的作用，需要进行全面的或“鞘脂组学”分析（对所有鞘脂类，或至少对关键亚类的所有成员进行结构特异性的定量分析），因为生物系统中的许多鞘脂类在结构和代谢上是相互转化的。尽管鞘脂类调节的细胞反应的多样性带来了实验挑战，但通过将高效液相色谱（HPLC）等经典分析分离技术与最先进的串联质谱（MS/MS）技术相结合，已经能够在单个样本中对多种鞘脂类进行检测和定量。作为脂质图谱联盟的一部分，开发了一种内标混合物，它包含信号代谢物（即鞘氨醇碱、鞘氨醇碱 - 1 - 磷酸酯、神经酰胺和神经酰胺 - 1 - 磷酸酯）以及更复杂的种类，如单 - 和二 - 己糖基神经酰胺和鞘磷脂。此外，随着更多内标变得可用，能够分析的种类数量正在迅速增加，包括脂肪酰基辅酶A、硫脂和其他复杂鞘脂类。由此产生的液相色谱 - 质谱/质谱分析是分析最严格的技术之一，能够以高通量为小样本量的“鞘脂组学”分析提供必要的灵敏度、结构特异性和定量精度。这篇综述总结了用于从游离鞘氨醇碱到更复杂的鞘脂类（如鞘磷脂、乳糖基神经酰胺和硫脂，包括那些目前被视为鞘脂类“第二信使”的中间产物）的鞘脂类鉴定、结构测定和定量的历史和最先进的分析技术。还讨论了一些新兴技术以及分析整个鞘脂组仍需解决的其他问题。
Sphingolipids comprise a highly diverse and complex class of molecules that serve as both structural components of cellular membranes and signaling molecules capable of eliciting apoptosis, differentiation, chemotaxis, and other responses in mammalian cells. Comprehensive or “sphingolipidomic” analyses (structure specific, quantitative analyses of all sphingolipids, or at least all members of a critical subset) are required in order to elucidate the role(s) of sphingolipids in a given biological context because so many of the sphingolipids in a biological system are inter-converted structurally and metabolically. Despite the experimental challenges posed by the diversity of sphingolipid-regulated cellular responses, the detection and quantitation of multiple sphingolipids in a single sample has been made possible by combining classical analytical separation techniques such as high-performance liquid chromatography (HPLC) with state-of-the-art tandem mass spectrometry (MS/MS) techniques. As part of the Lipid MAPS consortium an internal standard cocktail was developed that comprises the signaling metabolites (i.e. sphingoid bases, sphingoid base-1-phosphates, ceramides, and ceramide-1-phosphates) as well as more complex species such as mono- and di-hexosylceramides and sphingomyelin. Additionally, the number of species that can be analyzed is growing rapidly with the addition of fatty acyl Co-As, sulfatides, and other complex sphingolipids as more internal standards are becoming available. The resulting LC-MS/MS analyses are one of the most analytically rigorous technologies that can provide the necessary sensitivity, structural specificity, and quantitative precision with high-throughput for “sphingolipidomic” analyses in small sample quantities. This review summarizes historical and state-of-the-art analytical techniques used for the for the identification, structure determination, and quantitation of sphingolipids from free sphingoid bases through more complex sphingolipids such as sphingomyelins, lactosylceramides, and sulfatides including those intermediates currently considered sphingolipid “second messengers”. Also discussed are some emerging techniques and other issues remaining to be resolved for the analysis of the full sphingolipidome.
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影响因子：
3.1
作者：
Bergh, A;Bhattacharyya, S;Nilsson, UJ
通讯作者：
Nilsson, UJ
影响因子：
2.9
作者：
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DOI：
10.1016/j.bbalip.2006.02.021
发表时间：
2006-03-01
影响因子：
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作者：
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作者：
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