BRYOSTATIN BINDING SITE OF PKC AND BRYOSTATIN AN

PKC 和苔藓抑素 AN 的苔藓抑素结合位点

• 批准号: 2633921
• 负责人: JAMES A MORRISON
• 金额: $ 1.59万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2633921
• 关键词: X ray crystallography; active sites; allosteric site; bryostatin; chemical binding; chemical structure function; conformation; drug design /synthesis /production; enzyme inhibitors; enzyme structure; enzyme substrate analog; nuclear magnetic resonance spectroscopy; protein kinase C; radiotracer

The marine natural product, bryostatin, is in Phase II clinical trials in Britain for the treatment of cancer. The very low natural abundance of this class of compounds, a few parts per billion to several hundred parts per billion, and a lengthy synthesis of one member may hinder wider clinical studies. Bryostatins wide variety of biological effects is traced to binding to PKC(protein kinase C). The size and complexity of PKC precludes detailed investigation of its phorbol ester/diacylglycerol binding domain by X-ray crystallography or NMR. Smaller peptides (approximately 50 residues) which incorporate this subdomain can be studied by NMR. The binding of bryostatin to these peptides will be quantified using [26-3H]-bryostatin and qualitatively by NMR using 13C, 15N labeled peptides. As a medium resolution structure of one of these peptides is in hand, the determination of the exact structure of its bryostatin binding domain is a reasonable goal. Once the binding site is known, then the conformation of bryostatin in this site can be determined by 2-D NMR. Knowledge of this fit to the binding site will enable simplified analogs to be made which incorporate the features necessary for binding. Routes to potentially useful analogs are provided.

海洋天然产物苔藓抑素正在进行第二阶段临床试验， 英国治疗癌症。天然丰度极低的 这类化合物，十亿分之几到几百分之一 十亿分之一，一个成员的冗长合成可能会阻碍更广泛的 临床研究。 苔藓抑素的多种生物学效应可追溯到结合到 PKC（蛋白激酶C）。PKC的大小和复杂性排除了详细的 佛波酯/甘油二酯结合结构域X射线研究 晶体学或NMR。较小的肽（约50个残基）， 可以通过NMR研究引入该亚结构域。的结合 将使用[26- 3 H]-苔藓抑素定量苔藓抑素与这些肽的结合 并使用13 C、15 N标记的肽通过NMR进行定性。作为媒介 这些肽之一的解析结构在手， 确定其苔藓抑素结合结构域的确切结构， 合理的目标。一旦结合位点已知，那么构象 苔藓抑素在该位点的含量可通过2-D NMR测定。知道这起 与结合位点的配合将使得能够制备简化的类似物 结合装订所需的特征。潜在的路线 提供了有用的类似物。