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: 6394410
• 负责人: STEVEN E PFEIFFER
• 金额: $ 32.35万
• 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/6394410
• 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（来自申请人的摘要）：髓磷脂是一种动态的、功能性的 活性膜，对其环境（包括轴突）敏感。期间 髓磷脂生成过程中，它的产生量非常大。它是一个偏光 膜，具有宏观和微观结构域组织。它积极维护 其独特的结构以及与轴突的特殊关系。其中有趣的是 生化特性是高浓度的特定糖鞘脂， 特别是半乳糖脑苷脂（GalC）和脑硫脂。半乳糖脂具有 与少突胶质细胞分化的调节密切相关 和髓磷脂维护。 与胆固醇结合，目前的证据表明这些脂质 能够形成微域，其中一组离散的蛋白质成为 关联以实现通常与信号相关的专门功能 转导。虽然相当少量的“主要”髓磷脂蛋白 认识到，存在大量无法定量识别的“次要”问题 蛋白质。这两个类的功能尚不清楚。这个长期 该应用程序的目标是开发一个完整的数据库 髓磷脂蛋白髓磷脂蛋白质组，并开始关联的任务 与这些结构/代谢成分中的每一个一起发挥作用。 这将通过首先显示总髓磷脂蛋白来完成 补体作为二维凝胶电泳图，并识别 通过免疫标记、非埃德曼质谱法和 生物信息学。该信息将为该项目提供一个数据库 以及髓磷脂生物学界。其次，将进行详细分析 髓磷脂糖脂-胆固醇微结构域中的蛋白质， 将它们与总髓磷脂蛋白质组联系起来。将对他们进行一项研究 组织，并且将检验一个假设，该假设表明存在一个 可区分微域的马赛克。第三，该信息将用于 研究这些微域的功能生物学。小鼠模型在 不产生半乳糖脑苷脂和硫化物（CGT 无效小鼠） 将根据其微域进行分析。在这只鼠标中，节点 朗维尔被改变了；我们提出的是这些的分子相关性 形态变化，我们已经证明了一组离散的关联 具有糖脂微结构域的蛋白质发生了改变。最后，我们有 显示髓鞘少突胶质细胞糖蛋白（MOG）分为这些 微结构域，以及实验刺激其进入成熟 少突胶质细胞导致特定的酪氨酸快速磷酸化 蛋白质。这与抗 OMOG 介导的过敏的关系 会考虑脑脊髓炎。 预计从这些调查中得出的概念将有助于 更好地了解髓磷脂的生物发生、维护和正常功能，并帮助 设计范例以促进多种髓鞘质稳定性和髓鞘再生 硬化。