PDGF AA-DIMER STRUCTURE & ATHEROSCLEROSIS DRUG DESIGN

本项目将为PDGF-AA的结构分析奠定基础 第一阶段，导致其三维结构确定在第二阶段。 这项资助的最终目标是使用PDGF的3D结构来开发 抑制PDGF-AA在动脉粥样硬化中的作用的药物。这 方法将利用最现代的技术 生物技术和结构分析。 重组人将生产毫克量的PDGF AA-Homodimer DNA方法学；克隆的蛋白将在大肠杆菌中表达，随后 通过复性和最终纯化。具有生物活性的PDGF-AA 然后用来生长适合于单晶x-的同源二聚体晶体- 光线衍射研究。核磁共振光谱学将同时在 溶液中的蛋白质，为2D-核磁共振奠定基础 对其结构进行分析。适用于X射线衍射研究的晶体 高分辨率和/或核磁共振光谱数据适用于规划 详细的三维结构分析将构成第二阶段战略的基础。 使用计算算法加速建模的可行性将 也要接受评估。PDGF AA-Homodimer的详细3D结构如下 在第二阶段计划中确定，导致分子基础 合理设计阻断PDGF作用的药物在肿瘤发生发展中的作用 动脉粥样硬化斑块。

This project will develop the basis for structure analysis of PDGF-AA in Phase I, leading to its 3D structure determination in PHase II. The ultimate goal of this grant is to use the 3D structure of PDGF to develop drugs that inhibit the actions of PDGF-AA in atherosclerosis. This approach will take advantage of the most contemporary techniques in biotechnology and structure analysis. Milligram quantities of PDGF AA-homodimer will be produced by recombinant DNA methodology; the cloned protein will be expressed in E. coli, followed by its renaturation and final purification. Biologically active PDGF-AA will then be used to grow homodimer crystals suitable for single crystal x- ray diffraction studies. NMR spectroscopy will be pursued concurrently on the protein in solution, in order to establish the basis for 2D-NMR analysis of its structure. Crystals suitable for x-ray diffraction studies to high resolution and/or NMR spectral data suitable for planning a detailed 3D structure analysis would form the basis of a Phase II strategy. The feasibility of using computational algorithms to expedite modeling will also be assessed. A detailed 3D structure of PDGF AA-homodimer would be determine in the Phase II program, leading to a molecular basis for rational design of drugs that block PDGF action in the development of atherosclerotic plaque.