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脂质组学:通过电喷雾质谱法对细胞脂质进行分析。

Lipidomics: an analysis of cellular lipids by ESI-MS.

作者信息

Milne Stephen, Ivanova Pavlina, Forrester Jeffrey, Alex Brown H

机构信息

Department of Pharmacology and The Vanderbilt Institute of Chemical Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.

出版信息

Methods. 2006 Jun;39(2):92-103. doi: 10.1016/j.ymeth.2006.05.014.

Abstract

Recognition of the importance of lipid signaling in cellular function has led to rapid progress in the technology of lipid analysis. Measurements of lipid species changes are central to defining the networks of cell signaling (e.g., receptor activation by hormones or drugs) and lipids are involved in many biochemical and pathological processes. During the last several years our laboratory has focused on developing efficient methods for extraction of glycerophospholipids from biological systems and their detection and identification by mass spectrometry. We analyze phospholipid changes in mammalian cells as a result of a defined ligand stimulation strategy that supports the research questions of the consortium. The improvement of mass spectrometry techniques for phospholipid analysis combined with sophisticated computational methods developed in our group has facilitated simultaneous analysis of hundreds of phospholipid species in mammalian cells. This information is presented as Lipid Arrays (or more precisely as virtual arrays) and allows identification of temporal changes in membrane phospholipid species between two contrasting biological conditions (e.g., unstimulated basal vs. stimulated or as a contrast between normal and disease stages). Using the lipidomics approach, we are able to identify approximately 450 phospholipid species from total membrane extracts and qualitatively measure pattern response changes initiated by cell surface receptors. As such, this approach facilitates the elucidation of the metabolic changes induced by a perturbation in the cell and recognition of patterns of signaling.

摘要

对脂质信号在细胞功能中重要性的认识推动了脂质分析技术的迅速发展。脂质种类变化的测量是定义细胞信号网络(如激素或药物激活受体)的核心,并且脂质参与许多生化和病理过程。在过去几年中,我们实验室专注于开发从生物系统中提取甘油磷脂的有效方法,以及通过质谱对其进行检测和鉴定。我们通过一种明确的配体刺激策略来分析哺乳动物细胞中的磷脂变化,该策略支持联盟的研究问题。用于磷脂分析的质谱技术的改进,结合我们团队开发的复杂计算方法,促进了对哺乳动物细胞中数百种磷脂种类的同时分析。这些信息以脂质阵列(或更准确地说是虚拟阵列)的形式呈现,能够识别两种不同生物学状态(如未刺激的基础状态与刺激状态,或正常与疾病阶段之间的对比)下膜磷脂种类的时间变化。使用脂质组学方法,我们能够从总膜提取物中鉴定出约450种磷脂种类,并定性测量由细胞表面受体引发的模式反应变化。因此,这种方法有助于阐明细胞扰动引起的代谢变化以及信号模式的识别。

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