INTRACELLULAR TRANSLOCATION AND METABOLISM OF LIPIDS

脂质的细胞内易位和代谢

• 批准号: 2378190
• 负责人: RICHARD E PAGANO
• 金额: $ 32.06万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-03-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2378190
• 关键词: Golgi apparatus; affinity labeling; autoradiography; cell differentiation; cell growth regulation; ceramides; cerebrosides; digital imaging; endoplasmic reticulum; fluorescence microscopy; fluorescence spectrometry; intracellular membranes; laboratory rat; lipid bilayer membrane; lipid metabolism; lipid transport; liposomes; liver cells; membrane activity; membrane lipids; membrane reconstitution /synthesis; mutant; phospholipids; radiotracer; tissue /cell culture

The molecular mechanisms by which lipid molecules are synthesized, sorted, and transported to various cytoplasmic compartments within animal cells will be studied. A series of fluorescent lipid analogs will be used to correlate the intracellular distribution of lipid metabolites in living cells, as determined by fluorescence microscopy, with traditional lipid biochemical analyses. Where possible, similar studies will be performed using radiolabeled lipids and their precursors. The goals are to: (i) examine the role of vesicular and non- vesicular mechanisms of lipid transport to and from the plasma membrane; (ii) study the metabolism, sorting, and transport of sphingolipids through the ER/Golgi complex; (iii) study the metabolism and transport of a series of fluorescent and radioactive gangliosides in normal and mutant human skin fibroblasts; (iv) examine the sites of synthesis of some sphingolipids and their precursors in subcellular membrane fractions isolated from rat liver; and (v) study the interactions of liposomes with the intracellular membranes of semi-intact ("perforated") cells. Studies of the fluorescent lipids in living cells will be facilitated by resonance energy transfer microscopy, as recently developed in this laboratory. and by a recently acquired low-light-level digital imaging fluorescence microscope. Some studies will rely on the synthesis of a number of lipid analogs (and their stereoisomers) bearing fluorescent. radioactive, or photoaffinity labels. These will be used to distinguish various transport mechanisms from one another and to help identify the cellular proteins involved in lipid synthesis, sorting, and transport. The studies proposed in this application are basic to understanding membrane assembly and regulation of membrane lipid composition in cells and are fundamental to the development of rational treatments of membrane or cell surface- related disease states.

脂质分子的分子机制是 合成，分类和运输到各种细胞质 将研究动物细胞内的隔室。 一系列 荧光脂质类似物将用于将 活细胞中脂质代谢物的细胞内分布， 由荧光显微镜确定，传统脂质 生化分析。 在可能的情况下，类似的研究将是 使用放射性标记的脂质及其前体进行。 这 目标是：（i）检查囊泡和非 - 脂质转运的水泡机制往返等离子体 膜; （ii）研究代谢，分类和运输的 通过ER/高尔基体复合体的鞘脂； （iii）研究 一系列荧光和运输的代谢和运输 正常和突变人类皮肤中的放射性神经节 成纤维细胞； （iv）检查某些合成的位点 细胞膜中的鞘脂及其前体 从大鼠肝脏分离的馏分； （v）研究相互作用 半图的细胞内膜的脂质体 （“穿孔”）细胞。 研究荧光脂质中的 共振能量传递将促进活细胞 显微镜，该实验室最近开发的显微镜。 并通过 最近获得的低光级数字成像荧光 显微镜。 一些研究将依靠数量的综合 脂质类似物（及其立体异构体）带有荧光的含量。 放射性或光性标签。 这些将习惯 区分各种运输机制 有助于识别参与脂质合成的细胞蛋白， 分类和运输。 该应用中提出的研究 是理解膜组装和调节的基础 细胞中的膜脂质组成，对 膜或细胞表面合理处理的发展 相关疾病状态。