SYNTHETIC INHIBITORS OF MEMBRANE ANCHOR PHOSPHOLIPASES

膜锚定磷脂酶的合成抑制剂

• 批准号: 3306579
• 负责人: TSUNG-YING SHEN
• 金额: $ 11.72万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3306579
• 关键词: active sites; biological signal transduction; cell free system; chemical models; drug design /synthesis /production; enzyme substrate analog; glycolipids; membrane proteins; nuclear magnetic resonance spectroscopy; parasitic disease chemotherapy; phosphatidylinositols; phospholipase C; phospholipase D; phospholipase inhibitor

In the past four years increasing attention has been focused on a distinct type of membrane anchors of cell surface proteins, the glycosyl phosphatidylinositols (GPI). Among more than 40 of GPI-anchored proteins identified to date are various cell surface antigens, tumor markers, receptor systems, complement regulatory factors, cell adhesion molecules and membrane enzymes. The biosynthesis, metabolism and biological functions of glycolipid anchors are under active investigation. The release of surface proteins and GPI components with putative signal transduction properties by GPI-specific phospholipase C (GPI-PLC) and D (GPI-PLD) have been described in several parasitic and mammalian cell systems. Thus, there is a timely need for specific inhibitors and biochemical probes of these two phospholipases to facilitate the biological study of many important GPI-anchored proteins. Our preliminary data have shown that a unique GPI component previously synthesized in our laboratory selectively inhibit the trypanosome GPI- PLC, but not a PI-PLC, in a cell-free system at 1 mM in a stereospecific manner. We propose to use derivatives of this component as core structures to maintain GPI-specificity and develop more potent inhibitors of both GPI-PLC and a mammalian GPI-PLD. Various substituted, noncleavable and transition state analogs of the respective substrates will be synthesized. The optimal conformation of active inhibitors will be analyzed by NMR, X-ray and computer modeling. Related photoactivable biochemical probes will be prepared for a collaborator to characterize the active site of these enzymes. The effect of specific inhibitors on (1) the biosynthesis and metabolism of GPI-anchored proteins in Trypanosome brucei and Entamoeba histolytica, and (2) the signal transduction process of interleukins in human lymphocytes will also be investigated with collaborators towards the development of potential antiparasitic agents and novel immunomodulators.

在过去的四年里，越来越多的注意力集中在一个 细胞表面蛋白的不同类型的膜锚，糖基 磷脂酰肌醇（GPI）。 在40多种GPI锚定蛋白中， 迄今为止鉴定的是各种细胞表面抗原，肿瘤标志物， 受体系统，补体调节因子，细胞粘附分子 和膜酶。 生物合成、代谢和生物 糖脂锚的功能正在积极研究中。 的 释放表面蛋白质和GPI组分，具有推定的信号 GPI特异性磷脂酶C（GPI-PLC）和D的转导特性 GPI-PLD已在几种寄生虫和哺乳动物细胞中被描述 系统. 因此，及时需要特异性抑制剂， 这两种磷脂酶的生化探针，以促进 许多重要的GPI锚定蛋白的生物学研究。 我们 初步数据显示，一个独特的GPI组件以前 在我们实验室合成的选择性抑制锥虫GPI- PLC，但不是PI-PLC，在无细胞系统中以ImM在立体特异性微球中， 方式 我们建议使用该组件的衍生物作为核心 维持GPI特异性和开发更有效抑制剂的结构 GPI-PLC和哺乳动物的GPI-PLD。 各种取代， 各自底物的不可裂解的和过渡态类似物 将被合成。 活性抑制剂的最佳构象将 通过核磁共振、X射线和计算机建模进行分析。 相关光敏 将为合作者准备生化探针，以表征 这些酶的活性部位。 特异性抑制剂对 (1)GPI锚定蛋白的生物合成和代谢 布氏锥虫和溶组织内阿米巴;（2）信号 还将研究白细胞介素在人淋巴细胞中的转导过程。 与合作者一起进行调查，以开发潜力 抗寄生虫剂和新型免疫调节剂。