Structure-Function Study of Angiogenic Protein,Ephrin B2

血管生成蛋白Ephrin B2的结构与功能研究

• 批准号: 6752868
• 负责人: WALTER STAFFORD
• 金额: $ 31.53万
• 依托单位: BOSTON BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6752868
• 关键词: X ray crystallography; angiogenesis; computer simulation; crystallization; ephrins; fungal genetics; intermolecular interaction; laboratory mouse; membrane proteins; model design /development; molecular dynamics; molecular site; monoclonal antibody; peptide chemical synthesis; physical model; polymerization; protein structure function; protein tyrosine kinase; receptor; receptor binding; receptor expression; site directed mutagenesis; structural biology; thermodynamics; yeasts

The ephrins are membrane-bound proteins that act as ligands for the Eph Family of receptor tyrosine kinases. Ephrins and Eph receptors are involved in angiogenesis, axon guidance and topographic map formation, and segmentation. Evidence suggest that ephrin B2 and its receptor Eph B4, are the first molecular markers that distinguish arteries from veins, respectively, and these molecules are involved in the earliest stages of angiogenic remodeling of the primary capillary network established by vasculogenesis. Our long term objective is to obtain the structural basis for the function of the ephrins and the Eph receptors using X-ray crystallography. This information is critical for understanding the principles of angiogenesis and the relevant diseases such as cancer, eye diseases, and ischemia. Current studies on ephrins and Eph receptors focus on two key questions: 1) How ephrin extracellular domains cluster to activate Eph receptors. 2) What is the basis of the promiscuity of binding between Eph receptors and ephrins that is observed in vitro, which suggest a range of specificity that must be present in vivo. To address theses problems we propose to solve the X- ray crystal structure of ephrin B2 extracellular domain, for which we have already obtained well-diffracting crystals. We expect to determine the nature of ligand oligomerization, and see how the ligand might be bound by Eph receptors. The high- resolution, three-dimensional structure of ephrin B2 will allow us to design epitopes for monoclonal antibodies, and allow future directed-drug design, both of which could have therapeutic applications. Further, we propose to address the structural basis for Eph-ephrin binding by crystallizing and solving X-ray crystal structures of complexes of ephrin-Eph ligand binding domains. Finally, we propose to investigate the relationship between structure and function of the entire Eph receptor ectodomains via crystallography, to ascertain the function of the non-ligand binding domains.

肝配蛋白是作为受体酪氨酸激酶的Eph家族的配体的膜结合蛋白。 Ephrin和Eph受体参与血管生成、轴突引导和地形图形成以及分割。 有证据表明，肝配蛋白B2及其受体Eph B4分别是区分动脉和静脉的第一个分子标志物，并且这些分子参与由血管生成建立的初级毛细血管网络的血管生成重塑的最早阶段。 我们的长期目标是利用X射线晶体学获得ephrin和Eph受体功能的结构基础。 这些信息对于理解血管生成的原理和相关疾病如癌症、眼病和缺血至关重要。目前对ephrin和Eph受体的研究集中在两个关键问题上：1）ephrin胞外结构域如何簇合以激活Eph受体。 2)什么是在体外观察到的Eph受体和肝配蛋白之间结合的混杂性的基础，这表明一定存在于体内的特异性范围。 为了解决这些问题，我们提出解决肝配蛋白B2胞外结构域的X射线晶体结构，我们已经获得了衍射良好的晶体。 我们期望确定配体寡聚化的性质，并了解配体如何与Eph受体结合。 肝配蛋白B2的高分辨率三维结构将使我们能够设计单克隆抗体的表位，并允许未来的定向药物设计，这两者都可能具有治疗应用。 此外，我们建议通过结晶和解决肝配蛋白-Eph配体结合域的复合物的X射线晶体结构来解决Eph-ephrin结合的结构基础。 最后，我们建议通过晶体学研究整个Eph受体胞外结构域的结构与功能之间的关系，以确定非配体结合结构域的功能。