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De Novo Lipid Labeling for Comprehensive Analysis of Subcellular Distribution and Trafficking in Live Cells
bioRxiv - Cell Biology Pub Date : 2020-10-16 , DOI: 10.1101/2020.10.16.342683 Jun Zhang , Jia Nie , Haoran Sun , John-Paul Andersen , Yuguang Shi
bioRxiv - Cell Biology Pub Date : 2020-10-16 , DOI: 10.1101/2020.10.16.342683 Jun Zhang , Jia Nie , Haoran Sun , John-Paul Andersen , Yuguang Shi
Lipids exert dynamic biological functions which are determined both by their fatty acyl compositions and precise spatiotemporal distributions inside the cell. There are more than 1000 lipid species in a typical mammalian cell. However, it remains a daunting task to investigate any of these features in live cells for each of the more than 1000 lipid species. Here we resolved this issue by developing a de novo lipid labeling method for major lipid species, including glycerolipids, glycerophospholipids, and cholesterol esters by using a single fluorescent probe. The method not only allowed us to probe the precise subcellular distribution and trafficking of individual lipid species in live cells, but also uncovered some unexpected biological functions of previously reported lipid metabolic enzymes that were not possible by conventional biochemical methods. We envision that this method will become an indispensable tool for the functional analysis of individual lipid species and numerous lipid metabolic enzymes and transporters in live cells.
中文翻译:
De Novo脂质标记用于活细胞亚细胞分布和运输的综合分析
脂质发挥动态生物学功能,这既取决于它们的脂肪酰基组成,又取决于细胞内精确的时空分布。典型的哺乳动物细胞中有1000多种脂质。然而,对于超过1000种脂质种类的每一种,研究活细胞中的任何这些特征仍然是一项艰巨的任务。在这里,我们通过使用单个荧光探针开发一种主要脂质种类(包括甘油脂,甘油磷脂和胆固醇酯)的从头脂质标记方法来解决此问题。该方法不仅使我们能够探查活细胞中单个脂质物种的精确亚细胞分布和运输,而且还发现了以前报道的脂质代谢酶的某些意想不到的生物学功能,这是常规生化方法无法实现的。我们设想该方法将成为功能分析单个脂质种类和活细胞中众多脂质代谢酶和转运蛋白的必不可少的工具。
更新日期:2020-10-17
中文翻译:
De Novo脂质标记用于活细胞亚细胞分布和运输的综合分析
脂质发挥动态生物学功能,这既取决于它们的脂肪酰基组成,又取决于细胞内精确的时空分布。典型的哺乳动物细胞中有1000多种脂质。然而,对于超过1000种脂质种类的每一种,研究活细胞中的任何这些特征仍然是一项艰巨的任务。在这里,我们通过使用单个荧光探针开发一种主要脂质种类(包括甘油脂,甘油磷脂和胆固醇酯)的从头脂质标记方法来解决此问题。该方法不仅使我们能够探查活细胞中单个脂质物种的精确亚细胞分布和运输,而且还发现了以前报道的脂质代谢酶的某些意想不到的生物学功能,这是常规生化方法无法实现的。我们设想该方法将成为功能分析单个脂质种类和活细胞中众多脂质代谢酶和转运蛋白的必不可少的工具。