PDGF AA-DIMER STRUCTURE & ATHEROSCLEROSIS DRUG DESIGN

本研究将为PDGF-AA的结构分析奠定基础， 阶段I，导致其在阶段II中的3D结构确定。 的 这项资助的最终目标是利用PDGF的3D结构来开发 抑制PDGF-AA在动脉粥样硬化中作用的药物。 这 方法将利用最现代的技术， 生物技术和结构分析。 将通过重组质粒产生毫克量的PDGF AA-同源二聚体。 DNA方法，克隆的蛋白质将在E.大肠杆菌，随后 通过其复性和最终纯化。 生物活性PDGF-AA 然后将用于生长适合于单晶X-的同二聚体晶体。 射线衍射研究。 核磁共振光谱学将同时进行 溶液中的蛋白质，以建立2D-NMR的基础 分析其结构。 适用于X射线衍射研究的晶体 高分辨率和/或NMR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。 使用计算算法加快建模的可行性将 也要评估。 PDGF AA-同二聚体的详细3D结构将是 在第二阶段计划中确定，导致分子基础 合理设计药物，阻断血小板源生长因子的作用， 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.