| Using 14C-acetate Pulse-chase Labeling to Study Fatty Acid and Glycerolipid Metabolism in Plant Leaves
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Author:
Date:
2021-02-05
[Abstract] Lipids metabolism is comprised of networks of reactions occurred in different subcellular compartments. Isotopic labeling is a good way to track the transformations and movements of metabolites without perturbing overall cellular metabolism. Fatty acids, the building blocks of membrane lipids and storage triacylglycerols, are synthesized in plastids. The immediate precursor for fatty acid synthesis is acetyl-CoA. Exogenous acetate is rapidly incorporated into fatty acids in leaves and isolated plastids because it can diffuse freely through cellular membranes, enter the plastid where it is rapidly metabolized to acetyl-CoA. Therefore, isotope-labeled acetate is often used as a tracer for the investigation of fatty acid synthesis and complex lipid metabolism in plants and other organisms. ...
[摘要] [摘要]脂质代谢由发生在不同亚细胞区室的反应网络组成。同位素标记是跟踪代谢物的转化和运动的好方法，而不会干扰整个细胞的新陈代谢。发TTY酸，膜脂和存储的构建块的三酰基甘油，在质体中合成的。脂肪酸合成的直接前体是乙酰辅酶A。外源乙酸盐可快速掺入叶片和分离的质体中的脂肪酸中，因为它可以通过细胞膜自由扩散，进入质体，然后迅速代谢成乙酰辅酶A。 因此，同位素标记的乙酸盐通常用作研究植物和其他生物中脂肪酸合成和复杂脂质代谢的示踪剂。同位素标记的基本原理及其最新技术进展已得到综述（Allen等，2015）。本协议描述了使用 的14 C标记的乙酸，以确定的脂肪酸合成和降解速率和跟踪的代谢甘油脂中的叶子。该方法通常被称为醋酸酯脉冲追踪标记法，已被广泛用于探查脂质代谢的各个方面（Allen等，2015），包括自噬在膜脂质更新中的作用（Fan等，2015）。，2019）和脂质与淀粉代谢途径之间的相互作用（Yu et al。，2018）。
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| Extraction and Profiling of Plant Polar Glycerol Lipids
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Author:
Date:
2016-06-20
[Abstract] This protocol describes a method to extract total polar glycerol lipids from plant materials, followed by mass spectrometry profiling. Different glycerol lipid classes can be distinguished by their head-groups, which can be profiled automatically and quantitatively by a triple quadrupole mass spectrometry in multiple reaction monitoring (MRM) mode with an autosampler. Comparing with other established methods, such as thin layer chromatography (TLC) separation followed by Gas spectrometry (GC) analysis, this method requires little effort in sample preparation and separation, while the resolution is not limited to general lipid classes but at side chain level. This method was described and used successfully to profile plant lipids changes under freezing stress in Welti et al. ...
[摘要] 该方案描述了从植物材料提取总极性甘油脂质的方法，随后是质谱分析。 不同的甘油脂类可以通过其头基来区分，其可以通过使用自动进样器的多重反应监测（MRM）模式中的三重四极杆质谱自动和定量地进行概况分析。 与其他已建立的方法，例如薄层色谱（TLC）分离，然后气体光谱（GC）分析相比，该方法在样品制备和分离中几乎不需要付出努力，而分辨率不限于一般脂质类别，而是侧链水平 。 在Welti等人（2002）中描述并且成功地使用该方法来分析在冰冻胁迫下植物脂质的变化。
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| Positional Analysis of Fatty Acids in Phospholipids by PLA2 Treatment
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Author:
Date:
2016-06-20
[Abstract] Plant phospholipids can be produced in the endoplasmic reticulum or plastids. Lipids from different sources can be distinguished by the fatty acid profile, in terms of the preferred fatty acid species esterified to the sn-1 or sn-2 position of the glycerol backbone (Ohlrogge and Browse, 1995). This protocol is used to determine the fatty acid profile in total plant phospholipids by the treatment of sn-2 specific phospholipase A2 (PLA2).
[摘要] 植物磷脂可以在内质网或质体中产生。 来自不同来源的脂质可以通过脂肪酸谱来区分，根据酯化为甘油主链的sn-1或em-sn-2位置的优选脂肪酸种类 （Ohlrogge和Browse，1995）。 该方案用于通过处理sn-2特异性磷脂酶A 2（PLA 2）来确定总植物磷脂中的脂肪酸谱。 。
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