Three-in-One Simultaneous Extraction of Proteins, Metabolites and Lipids for Multi-Omics.
Three-in-One Simultaneous Extraction of Proteins, Metabolites and Lipids for Multi-Omics.
复制标题
DOI：
10.3389/fgene.2021.635971
复制
发表时间：
2021
影响因子：
3.7
通讯作者：
Chen S
中科院分区：
文献类型：
--
作者：
Kang J;David L;Li Y;Cang J;Chen S
阐明复杂的分子网络需要综合分析分子特征和不同水平的信息流和调节的变化。因此，高通量功能基因组学工具如转录组学、蛋白质组学、代谢组学和脂质组学已经出现以提供系统范围的研究。不幸的是，不同类型的生物分子的分析需要特定的样品提取程序与特定的分析仪器相结合。最有效的提取方案往往只涵盖有限类型的生物分子，由于它们不同的物理化学性质。因此，需要几组/等份样品用于提取不同的分子。在这里，我们采用了一种双相分馏方法，从同一样品中提取蛋白质，代谢物和脂质（3合1），用于液相色谱-串联质谱（LC-MS/MS）多组学。为了证明改进方法的实用性，我们使用细菌引发的拟南芥叶从同一样品中生成多组学数据集。总的来说，我们能够在单个样品中分析1849种蛋白质，1967种代谢物和424种脂质。这些分子涵盖了广泛的生物和分子过程，并允许对不同的分子和途径进行定量分析。我们的研究结果显示了多组学方法的明显优势，包括样品保存，高重现性和不同类型生物分子之间的紧密相关性。
Elucidation of complex molecular networks requires integrative analysis of molecular features and changes at different levels of information flow and regulation. Accordingly, high throughput functional genomics tools such as transcriptomics, proteomics, metabolomics, and lipidomics have emerged to provide system-wide investigations. Unfortunately, analysis of different types of biomolecules requires specific sample extraction procedures in combination with specific analytical instrumentation. The most efficient extraction protocols often only cover a restricted type of biomolecules due to their different physicochemical properties. Therefore, several sets/aliquots of samples are needed for extracting different molecules. Here we adapted a biphasic fractionation method to extract proteins, metabolites, and lipids from the same sample (3-in-1) for liquid chromatography-tandem mass spectrometry (LC-MS/MS) multi-omics. To demonstrate utility of the improved method, we used bacteria-primed Arabidopsis leaves to generate multi-omics datasets from the same sample. In total, we were able to analyze 1849 proteins, 1967 metabolites, and 424 lipid species in single samples. The molecules cover a wide range of biological and molecular processes, and allow quantitative analyses of different molecules and pathways. Our results have shown the clear advantages of the multi-omics method, including sample conservation, high reproducibility, and tight correlation between different types of biomolecules.