PROTEOMIC MAPPING OF MYELIN AND ITS MEMBRANE SUBDOMAINS

髓磷脂及其膜亚域的蛋白质组图谱

• 批准号: 6529674
• 负责人: STEVEN E PFEIFFER
• 金额: $ 32.54万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6529674
• 关键词: biotechnology; cholesterol; combinatorial chemistry; functional /structural genomics; gel electrophoresis; glycosphingolipids; informatics; laboratory mouse; mass spectrometry; molecular biology information system; myelin; nerve /myelin protein; node of Ranvier; protein structure function

DESCRIPTION (From the Applicant's Abstract):Myelin is a dynamic, functionally active membrane, sensitive to its environment including axons. During myelinogenesis, it is produced in prodigious quantities. It is a polarized membrane, with both macro- and microdomain organization. It actively maintains its unique structure and special relationship with axons. Among its interesting biochemical properties is a high concentration of specific glycosphingolipids, in particular galactocerebroside (GalC) and sulfatide. The galactolipids have been strongly implicated in the regulation of oligodendrocyte differentiation and myelin maintenance. In combination with cholesterol, current evidence indicates that these lipids are able to form microdomains in which a discrete set of proteins become associated in order to effect specialized functions often related to signal transduction. While a rather small number of "major" myelin proteins are recognized, there is a plethora of poorly identified quantitatively "minor" proteins. The function of both classes is poorly understood. This long term goal of this application is to develop a data base of the total complement of myelin proteins the Myelin Proteome, and to begin the task of associating function with each of these structural/metabolic components. This will be accomplished by first displaying the total myelin protein complement as a two-dimensional gel electrophoretic map, and identifying the individual members by immunolabeling, non-Edman mass spectrometry, and bioinformatics. This information will provide a database both for this project and for the myelin biology community. Second, a detailed analysis will be made of the proteins in the glycosphinogolipid-cholesterol microdomains of myelin, relating them to the total myelin proteome. A study will be made of their organization, and a hypothesis will be tested that proposes that there is a mosaic of distinguishable microdomains. Third, this information will be used to investigate the functional biology of these microdomains. A mouse model in which galactosylcerebroside and sulfide are not produced (the CGT-null mouse) will be analyzed in terms of its microdomains. In this mouse the nodes of Ranvier are altered; in what we propose are the molecular correlates of these morphological changes, we have shown that the association of a discrete set of proteins with the glycosphinogolipid microdomains is altered. Finally, we have shown that myelin-oligodendrocyte glycoprotein (MOG) partitions into these microdomains, and the experimental stimulation its entry in maturing oligodendrocytes results in the rapid tyrosine phosphorylation of specific proteins. The relationship of this to antiOMOG mediated allergic encephalomyelitis will be considered. It is expected that concepts derived from these investigations will help to better understand myelin biogenesis, maintenance and normal function, and help to design paradigms to promote myelin stability and remyelination in multiple sclerosis.

描述（摘自申请人的摘要）：髓磷脂是一种动态的功能 活性膜，对包括轴突在内的环境敏感。期间 骨髓生成，它以大量数量产生。这是两极分化的 膜，具有宏观和微区组织。它积极维持 它与轴突的独特结构和特殊关系。在它的有趣之中 生化特性是高浓度的特定糖磷脂， 尤其是半乳脑溴化物（GALC）和亚硫酸盐。半乳脂具有 与少突胶质细胞分化的调节密切相关 和髓鞘维护。 结合胆固醇，当前的证据表明这些脂质 能够形成一个微区，其中一组离散的蛋白质变成 关联以实现通常与信号有关的专业功能 转导。虽然少数“主要”髓磷脂蛋白是 公认的是，有很多数量的“次要” 蛋白质。两个类别的功能都鲜为人知。这长期 该应用的目标是建立总补充的数据库 髓磷脂蛋白髓磷脂蛋白质组，并开始关联的任务 这些结构/代谢成分中的每一个。 这将通过首先显示总髓磷脂蛋白来完成 作为二维凝胶电泳图的补充，并识别 通过免疫标签，非埃德曼质谱法和个人成员 生物信息学。此信息将为该项目提供两个数据库 以及髓磷脂生物学社区。其次，将进行详细的分析 髓磷脂的糖磷脂 - 胆固醇微域中的蛋白质， 将它们与总髓磷脂蛋白质组联系起来。将对他们的研究进行研究 组织，并将检验一个假设，该假设提出有一个 可区分微区的镶嵌物。第三，此信息将用于 研究这些微区的功能生物学。鼠标模型 未产生哪种半乳核苷和硫化物（CGT-NULL小鼠） 将根据其微区域进行分析。在这只鼠标中 兰维尔被改变；我们提出的是这些分子相关性 形态学变化，我们已经证明了一组离散集的关联 具有糖磷脂微域的蛋白质改变了。最后，我们有 表明髓磷脂 - 寡糖细胞糖蛋白（MOG）分区 微域以及实验刺激其在成熟中的进入 少突胶质细胞导致特异性酪氨酸磷酸化的快速磷酸化 蛋白质。这与抗瘤介导的过敏性的关系 将考虑脑脊髓炎。 预计从这些调查中得出的概念将有助于 更好地了解髓磷脂生物发生，维护和正常功能，并有助于 设计范式以促进多种 硬化。