Stable lsotope-Assisted Lipidomics Combined with Nontargeted lsotopomer Filtering, a Tool to Unravel the Complex Dynamics of Lipid Metabolism
Stable lsotope-Assisted Lipidomics Combined with Nontargeted lsotopomer Filtering, a Tool to Unravel the Complex Dynamics of Lipid Metabolism
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稳定同位素辅助脂质组学与非靶向同位素过滤相结合，是揭示脂质代谢复杂动力学的工具
DOI：
10.1021/ac400293y
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发表时间：
2013-05-07
影响因子：
7.4
通讯作者：
Xu, Guowang
中科院分区：
文献类型：
--
作者：
Li, Jia;Hoene, Miriam;Xu, Guowang
复杂的代谢机制和网络的调查已成为后基因组领域的研究重点，从而创造了复杂的分析方法的需求日益增加。一种这样的工具是脂质组学分析，其提供在给定时间系统的脂质组成的详细图片。将稳定同位素引入所研究的系统中可以另外提供关于单个脂质种类的合成、转化和降解的信息。然而，捕获脂质网络的整个动态仍然是一个挑战。我们开发并评估了一种新的策略，用于深入分析脂质代谢的动态，具有高分子特异性和网络覆盖的能力。一般工作流程包括稳定的同位素标记实验，超高效液相色谱（UHPLC）/高分辨率轨道质谱（MS）脂质分析和数据处理的软件工具，用于全球同位素异构体过滤和匹配。作为概念的证明，这种方法被应用于与[U-C-13]-棕榈酸盐一起培养4、12和24小时的原代人骨骼肌细胞中动态脂质代谢的网络范围映射。在肌细胞脂质提取物中，检测到692种同位素异构体，可分配给203种标记的脂质种类，跨越12个脂质（亚）类。有趣的是，一些脂质类显示出高周转率，但稳定的总量，而其他的棕榈酸酯治疗过程中的量增加。这里提出的新策略有可能打开新的详细了解脂质代谢的动态，可能会导致更好地了解生理机制和代谢扰动。
Investigations of complex metabolic mechanisms and networks have become a focus of research in the postgenomic area, thereby creating an increasing demand for sophisticated analytical approaches. One such tool is lipidomics analysis that provides, a detailed picture of the lipid composition of a system at a given time Introducing stable isotopes into the studied system can additionally provide information on the synthesis, transformation and degradation of individual lipid species. However, capturing the entire dynamics of lipid networks is still a challenge. We developed and evaluated a novel strategy for the in-depth analysis of the dynamics of lipid metabolism with the capacity for high molecular specificity and network coverage. The general workflow consists of stable isotope-labeling experiments, ultrahigh-performance liquid chromatography (UHPLC)/high-resolution Orbitrap-mass spectrometry (MS) lipid profiling and data processing by a software tool for global isotopomer filtering and matching. As a proof of concept, this approach was applied to the network-wide mapping of dynamic lipid metabolism in primary human skeletal muscle cells cultured for 4, 12, and 24 h with [U-C-13]-palmitate. In the myocellular lipid extracts, 692 isotopomers were detected that could be assigned to 203 labeled lipid species spanning 12 lipid (sub)classes. Interestingly, some lipid classes showed high turnover rates but stable total amounts while the amount of others increased in the course of palmitate treatment. The novel strategy presented here has the potential to open new detailed insights into the dynamics of lipid metabolism that may lead to a better understanding of physiological mechanisms and metabolic perturbations.