Lipid Dynamics in the Golgi Apparatus

高尔基体中的脂质动力学

基本信息

批准号：
10388874
负责人：
Christopher G Burd
金额：
$ 12.5万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2022-06-30
项目状态：
已结题

项目摘要

Project Summary Each organelle of the cell has a unique lipid and protein composition that confers its specific functions. While much has been learned regarding the mechanisms that mediate sorting of proteins to different organelles, the mechanisms that govern the lipid content of organelles are largely unknown. New lipid synthesis occurs in the endoplasmic reticulum (ER), the Golgi apparatus, and the mitochondrion, and they are then distributed to other organelles. There are expansive gaps in knowledge regarding the mechanisms that coordinate the synthesis and distributions of lipids. The research proposed in this project focuses on sphingomyelin, a major structural component of the cellular membranes, and the Golgi apparatus, which functions as a “lipid based sorting station” that disseminates lipids and proteins to nearly all organelles of the cell. Multiple types of cargo transport vesicles bud from the Golgi and it is widely thought that coalescence of lipids such as sphingolipids and cholesterol is a driving force for sorting of cargos into to transport pathways. Preliminary data document the engineering of a natural sphingomyelin-binding protein into a biosensor that enables analyses of sphingomyelin dynamics (synthesis, trafficking) in living cells. We will apply this new tool in fluorescence microscopy-based experiments to determine if sphingomyelin is enriched in distinct Golgi-to-plasma membrane transport carriers. Protein ‘clients’ of the sphingomyelin transport pathway will be identified using enzymatic proximity labeling and proteomic analyses, and sphingomyelin-dependent sorting mechanisms for these proteins will be elucidated. The research will provide new insights into the regulation and roles of sphingomyelin within the cell.

项目概要细胞的每个细胞器都有独特的脂质和蛋白质成分，赋予其特定的功能。关于介导蛋白质分类到不同细胞器的机制已经了解了很多控制细胞器脂质含量的机制在很大程度上是未知的。内质网（ER）、高尔基体和线粒体，然后它们被分配到其他关于协调合成的机制存在巨大的知识空白。该项目提出的研究重点是鞘磷脂，一种主要的结构。细胞膜的组成部分和高尔基体，其功能是“基于脂质的分类” “站”将脂质和蛋白质传播到细胞的几乎所有细胞器中的多种类型的货物。运输囊泡从高尔基体中萌芽，人们普遍认为，鞘脂等脂质的聚结胆固醇是将货物分类到运输路径的驱动力。将天然鞘磷脂结合蛋白工程化为生物传感器，从而能够分析活细胞中的鞘磷脂动力学（合成、运输）我们将在荧光中应用这种新工具。基于显微镜的实验，以确定鞘磷脂是否富含不同的高尔基体到质膜鞘磷脂运输途径的蛋白质“客户”将使用酶法进行鉴定。邻近标记和蛋白质组分析，以及鞘磷脂依赖性分选机制该研究将为蛋白质的调节和作用提供新的见解。细胞内的鞘磷脂。