In Search of Mannolipid Intermediate Flippases

寻找甘露糖脂中间翻转酶

基本信息

批准号：
8341035
负责人：
Charles J Waechter
金额：
$ 28.22万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-20 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8341035
关键词：
1,2-diacylglycerol Anabolism Benzophenones Biological Assay Cell membrane Cells Chemistry Collection Congenital Disorders Defect Diagnosis Diffuse Diglycerides Dolichol Dolichol Monophosphate Mannose Endoplasmic Reticulum Eukaryota Eukaryotic Cell Face GPI Membrane Anchors Genes Genetic Genomic Library Glycolipids Goals Human In Vitro Insulin Receptor Label Laboratories Lead Link Lipids Mammalian Cell Mannose Mediating Membrane Membrane Glycoproteins Membrane Proteins Metabolic Micrococcus luteus Movement Mutation Oligosaccharides Pathway interactions Play Polysaccharides Process Prokaryotic Cells Protein Biosynthesis Protein C Proteins Proteomics Protoplasts Research Design Rhodopsin Scheme Sodium Channel Suicide Surface Temperature Vesicle Yeasts analog experience extracellular gene discovery glycosylation interest lipomannan man monolayer mutant novel protein function seal temperature sensitive mutant voltage

项目摘要

DESCRIPTION (provided by applicant): During the past 35 years this laboratory has been studying the function of polyisoprenol-linked intermediates in glycan synthesis in prokaryotes and eukaryotes. A long-standing interest in the topological issues involved in the biosynthesis of extracellular saccharides has led us to focus on the proteins that mediate the transbilayer movements of the lipid-linked intermediates utilized as glycosyl donors in these pathways. The current proposal is aimed at identifying the proteins that mediate the transbilayer movement of dimannosyldiacylglycerol and mannosylphosphoryldolichol, two mannolipid intermediates in the assembly of the lipomannan of Micrococcus luteus. These studies will provide important clues to the identification of related proteins functioning in protein N-glycosylation, C- and O-mannosylation of proteins and biosynthesis of glycosylphosphatidylinositol anchors in humans. The discovery of genes encoding membrane proteins functioning as flippases in these assembly processes will be valuable in the diagnosis of potential genetic errors referred to as Congenital Disorders in Glycosylation (CDGs). PUBLIC HEALTH RELEVANCE: This application proposes studies designed to identify the membrane proteins (flippases) in Micrococcus luteus that mediate the transbilayer movements of dimannosyl-diacylglycerol and mannosylphosphorylundecaprenol, two mannolipid intermediates in bacterial lipomannan synthesis. These studies on micrococcal lipomannan assembly will provide important new clues to the identification of related membrane proteins functioning in protein N-glycosylation, C- and O-mannosylation of proteins, and the biosynthesis of glycosylphosphatidylinositol (GPI) anchors in humans. Since N-linked oligosaccharides play a key role in the proper folding and intracellular routing of important membrane glycoproteins, e.g. rhodopsin, voltage-sensitive sodium channels and the insulin receptor, the identification of the genes encoding mammalian flippases is relevant to the diagnosis of potential genetic errors referred to as Congenital Disorders in Glycosylation (CDGs).

描述（由申请人提供）：在过去的35年中，该实验室一直在研究原核生物和真核生物的聚糖中间体中多肾上腺素连接的中间体的功能。对细胞外糖的生物合成涉及的拓扑问题的长期兴趣使我们专注于介导这些途径中利用为糖基供体的脂质连接中间体的跨贝贝运动的蛋白质。当前的建议旨在鉴定介导Dimannosyldiacyl -Glycerol和Mannosylphopphopphosphosphordolichol的跨贝碱运动的蛋白质，这是小脂肪菌的Lipomannan组装中的两个甘露脂素中间体。这些研究将为鉴定蛋白质N-糖基化，C-和O-甘露糖基化的相关蛋白质的鉴定提供重要的线索，以及人类中糖基磷脂酰肌醇锚固的生物合成。在这些组装过程中编码膜蛋白的基因的发现对于诊断为糖基化（CDGS）中的潜在遗传误差（CDGS）的潜在遗传误差将是有价值的。公共卫生相关性：该应用程序提出的研究旨在鉴定小叶片中的膜蛋白（FLIPPase）介导二甘糖基 - 二酰基甘油和甘露磷酸磷酸氯糖糖醇的跨贝族运动，这是两种甘诺果磷酸甘油糖，两种甘诺果中间体中的两种甘诺果中间体。这些关于小局局部脂多曼南组装的研究将为鉴定蛋白质N-糖基化，蛋白质的C和O-甘露糖基化的相关膜蛋白的鉴定提供重要的新线索，并为蛋白质的c-和o-甘露素化以及糖基磷脂酰磷脂酰素素（GPI）锚固的糖基质基因合成。由于N连接的寡糖在重要的膜糖蛋白的适当折叠和细胞内路由中起着关键作用，例如视紫红质，电压敏感的钠通道和胰岛素受体，编码哺乳动物flippases的基因的鉴定与诊断为糖基化（CDGS）中的潜在遗传错误的诊断。