Structural aspects of urokinase receptor in vascular biology

血管生物学中尿激酶受体的结构方面

• 批准号: 7340491
• 负责人: Mingdong Huang
• 金额: $ 38.25万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-15 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7340491
• 关键词: Adhesives; Alzheimer' s Disease; Binding; Biological Process; Biology; Blood Vessels; Cell Adhesion; Cell Surface Receptors; Cellular biology; Chemotaxis; Classification; Coagulation Process; Complex; Cytolysis; Data Collection; Drosophila genus; Enzyme Precursors; Epitopes; Fab Immunoglobulins; Future; G Protein-Coupled Receptor Genes; Glycoproteins; Goals; HIV-1; Human; Infection; Inflammation; Integrins; Length; Ligands; Mediating; Methods; Modeling; Molecular; Monoclonal Antibodies; Multiple Sclerosis; Nature; Neoplasm Metastasis; Pathologic; Pathologic Processes; Peptides; Physiological; Plasminogen; Play; Pliability; Preparation; Prevention; Principal Investigator; Production; Protease Domain; Proteins; Publishing; Reagent; Recombinant Proteins; Recombinants; Research Personnel; Resolution; Roentgen Rays; Role; Science; Signal Transduction; Solid; Structure; Study Section; Surface; System; Techniques; Testing; Therapeutic; Tumor Cell Invasion; Urokinase; Urokinase Plasminogen Activator Receptor; Vascular Diseases; Vitronectin; X ray diffraction analysis; X-Ray Crystallography; X-Ray Diffraction; angiogenesis; base; cell motility; chemical synthesis; computerized data processing; conformational alteration; design; mutant; neointima formation; programs; protein expression; protein protein interaction; receptor; receptor binding; small molecule; stem; tool; tumor

DESCRIPTION (provided by applicant): Urokinase plasminogen activator (uPA) together with its cell surface receptor (uPAR) mediates surface-bound plasminogen activation and has been recognized to play essential roles in several biological processes important in vascular biology, including clot lysis, cell adhesion, cell migration, inflammation, signal transduction, angiogenesis, and chemotaxis. Therefore, the study of this system will be critical to understand the causes, prevention, and treatment of vascular disorders and diseases including tumor invasion, tumor metastasis, neointima formation, infection with human immunodeficiency virus-1 (HIV-1), Alzheimer disease, and multiple sclerosis. The molecular basis for these broad physiological and pathophysiological roles of uPAR/uPA system stems from uPAR's capability to interact with many diverse ligands, e.g., uPA, vitronectin, integrins (31, 32, 33), LRP, GPCR, and the dynamic conformational changes caused by these protein-protein interactions. The principal investigator's recent results (Huai, et al. "Structure of Human Urokinase Plasminogen Activator in Complex", Science, in press, 2006) suggest conformational flexibility of uPAR. The goal of this proposal is to systematically study the structural basis and the dynamic nature of the interactions between uPAR and its ligands. The principal investigator proposes to study the structures of soluble uPAR (suPAR) (aim 1), suPAR in complex with vitronectin (aim 2), suPAR in complex with full- length uPA (aim 3), and to compare and analyze the structures of suPAR and suPAR-ligand complexes (aim 4). Protein X-ray crystallography will be the main tool to study these protein-protein interactions, but other common molecular and cell biology techniques will also be used. The principal investigator has established methods to generate needed reagents and has made several preliminary suPAR-ligand crystals, providing a solid basis for these proposed studies. These systematic structural studies of uPAR and its complexes with physiologically important uPA forms and vitronectin will understand the dynamic interactions between uPAR and its ligands, identify unanimously the binding determinants on the receptor and the ligands that confer therapeutic accessibility, and provide a framework for future design and optimization of small molecular antagonists to regulate these protein-protein interactions and to intervene the pathologic consequences resulting from these interactions.

描述（由申请方提供）：尿激酶纤溶酶原激活剂（uPA）及其细胞表面受体（uPAR）介导表面结合纤溶酶原激活，并已被认为在血管生物学中的几个重要生物学过程中发挥重要作用，包括凝块溶解、细胞粘附、细胞迁移、炎症、信号转导、血管生成和趋化性。因此，对该系统的研究对于了解血管疾病和疾病的原因、预防和治疗至关重要，这些疾病包括肿瘤侵袭、肿瘤转移、新生内膜形成、人类免疫缺陷病毒-1（HIV-1）感染、阿尔茨海默病和多发性硬化。uPAR/uPA系统的这些广泛生理和病理生理作用的分子基础源于uPAR与许多不同配体相互作用的能力，例如，uPA、玻连蛋白、整联蛋白（31、32、33）、LRP、GPCR以及由这些蛋白质-蛋白质相互作用引起的动态构象变化。主要研究者最近的结果（Huai等人，“Structure of Human Urokinase Plasminogen Activator in Complex”，Science，in press，2006）表明uPAR的构象柔性。本研究的目的是系统地研究uPAR与其配体之间相互作用的结构基础和动力学性质。主要研究者建议研究可溶性uPAR（suPAR）（目的1）、suPAR与玻连蛋白复合物（目的2）、suPAR与全长uPA复合物（目的3）的结构，并比较和分析suPAR和suPAR-配体复合物的结构（目的4）。蛋白质X射线晶体学将是研究这些蛋白质-蛋白质相互作用的主要工具，但也将使用其他常见的分子和细胞生物学技术。主要研究者已经建立了生成所需试剂的方法，并制备了几种初步的suPAR-配体晶体，为这些拟议的研究提供了坚实的基础。这些uPAR及其与生理上重要的uPA形式和玻连蛋白的复合物的系统结构研究将理解uPAR及其配体之间的动态相互作用，一致地鉴定受体和配体上赋予治疗可及性的结合决定簇，并为未来设计和优化小分子拮抗剂以调节这些蛋白质提供了框架-蛋白质的相互作用，并干预这些相互作用导致的病理后果。