DESIGN OF CHEMICAL PROBES TO STUDY PHOSPHOLIPASE C

研究磷脂酶 C 的化学探针的设计

• 批准号: 3301616
• 负责人: STEPHEN MARTIN
• 金额: $ 11.48万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3301616
• 关键词: X ray crystallography; chemical synthesis; enzyme mechanism; enzyme substrate analog; isozymes; phosphatidylinositols; phospholipase C; phospholipid inhibitor; phospholipids; protein structure function; radiotracer

The overall goal of this research program is to design and develop chemical probes that may be exploited to enhance our understanding of the mechanism of action of bacterial and mammalian isoenzymes of the phospholipase C (PLC) class. Substrate analogues that may serve as potential mechanistic probes and inhibitors will be the synthetic targets of these efforts. Inasmuch as it is known that the mammalian phosphoinositide-specific PLC plays a key role signal transduction by releasing the second messengers 1,2-diacylglycerol (DAG) and 1,4,5-inositol trisphosphate (IP3), these studies should enable elucidation of certain mechanistic features of this important process. Compounds prepared during these studies could have beneficial impact in a variety of disease areas, including anticancer, cardiovascular, and anti-inflammatory. The principal foci of these investigations will be to: (1) develop efficient, general methods for the syntheses of all classes of phospholipids and derivatives thereof that contain modified head groups and/or modified phosphatidic acid subunits; (2) design and prepare rationally selected phospholipids to test hypotheses regarding the mechanism of enzymatic hydrolysis of the phosphodiester bond in different classes of phospholipids; (3) design and synthesize phospholipid substrate analogues for biological screening as potential inhibitors of bacterial and mammalian PLC isoenzymes; and (4) collaborate in single crystal X-ray studies of inhibitors complexed with bacterial PLC to examine phospholipid- enzyme interactions thereby gaining insights to design second generation mechanistic probes. Biological assays and screening experiments to survey structure-activity relationships with bacterial PLC (B. cereus) and PI-PLC (B. thuringiensis) will be executed in our laboratories according to standard protocols. The X-ray crystallographic studies of inhibitors of bacterial PLC will be carried out in collaboration with Professor Edward Hough (University of Tromso, Norway). The in vitro screening for structure-activity relationships with mammalian PI-PLC and PIP2-PLC will be performed in collaboration with scientists at DuPont (Dr. Pat N. Confalone's group), with Professor Philip J. Majerus (Washington University School of Medicine) and Dr. Sue Goo Rhee (National Institutes of Health, Heart, Lung and Blood Institute).

该研究计划的总体目标是设计和开发化学 可以用来增强我们对机制的理解的探针 磷脂酶C的细菌和哺乳动物同工酶的作用 (PLC)课 底物类似物可能作为潜在的机制 探针和抑制剂将是这些努力的合成目标。 由于已知哺乳动物磷酸肌醇特异性PLC 通过释放第二信使在信号转导中起关键作用 1，2-二酰基甘油（DAG）和1，4，5-三磷酸肌醇（IP 3），这些 研究应该能够阐明某些机制的特点， 重要的过程。 在这些研究期间制备的化合物可能具有 在多种疾病领域，包括抗癌， 心血管和抗炎。 这些调查的主要重点将是：（1）发展 有效的，一般的方法，用于合成的所有类别的 含有修饰的头部基团的磷脂及其衍生物 和/或修饰的磷脂酸亚基;（2）设计和制备 合理选择的磷脂，以测试关于 酶水解磷酸二酯键的机制，在不同的 磷脂的种类;（3）磷脂底物的设计与合成 用于生物筛选作为细菌和真菌的潜在抑制剂的类似物， 哺乳动物PLC同工酶;和（4）在单晶X射线 与细菌PLC复合的抑制剂的研究，以检查磷脂- 酶的相互作用，从而获得设计第二代 机械探测器 生物测定和筛选实验，以调查结构-活性 与细菌PLC（B. cereus）和PI-PLC（B.苏云金杆菌） 将在我们的实验室按照标准方案进行 的 细菌PLC抑制剂的X射线晶体学研究将是 与爱德华·霍夫教授（牛津大学）合作进行的 挪威特罗姆瑟）。 体外结构活性筛选 与哺乳动物PI-PLC和PIP 2-PLC的关系将在 与杜邦公司的科学家合作（Pat N. Confalone的小组）， Philip J. Majerus教授（华盛顿大学医学院） 和Sue Goo Rhee博士（美国国立卫生研究院，心脏，肺和血液 研究所）。