Regulation of von Willebrand Factor - platelet binding by Force and Interdomain I

通过力和域间 I 调节血管性血友病因子 - 血小板结合

• 批准号: 8197730
• 负责人: Wendy E Thomas
• 金额: $ 41.92万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8197730
• 关键词: Adhesions; Adhesives; Adverse effects; Antibodies; Behavior; Binding; Biological Assay; Blood Platelets; Blood Vessels; Cardiovascular Diseases; Cause of Death; Cells; Cloning; Disease; Elements; Event; Exposure to; Glycoprotein Ib; Hemorrhage; Hemostatic function; Knowledge; Lead; Measures; Mechanics; Mediating; Microfluidics; Mutation; Myocardial Infarction; Nucleic Acid Regulatory Sequences; Physiological; Physiology; Plasma Proteins; Platelet aggregation; Point Mutation; Process; Protein Engineering; Recombinants; Regulation; Research Proposals; Role; Scanning Probe Microscopes; Site; Spectrum Analysis; Stress; Stretching; Stroke; Surface; System; Testing; Therapeutic Agents; Thrombosis; Thrombus; Work; base; fluid flow; in vivo; novel; prevent; receptor; research study; shear stress; single molecule; theories; von Willebrand Disease; von Willebrand Factor

Project Summary Arterial thrombosis occurs in conditions of high fluid flow and is mediated by the binding of platelets via their receptor Glycoprotein Ib (GPIb) to the plasma protein called von Willebrand Factor (VWF) via the A1 domain. This process is normally down-regulated to prevent spontaneous adhesion and thrombosis, but is activated by high shear stress. The proposed work will test the hypothesis that binding of GPIb to A1 is down-regulated by the neighboring domains or flanking regions within VWF, so that VWF displays interdomain auto-inhibition. It will also test the hypothesis that mechanical force associated with high shear stress activates platelet binding by pulling these regulatory regions away from the A1 domain to relieve the auto- inhibition. The final hypothesis to be tested is that conditions that increase the spontaneous binding of platelets to VWF, including type 2B von Willebrand disease, do so by damaging the normal interdomain auto-inhibition. Regulatory domains and elements within VWF will be identified by cloning different regions of VWF and determining the adhesive behavior of both soluble and surface-immoblized molecules. The effect of mechanical force and shear stress on this regulation will be determined using an atomic force microscope and microfluidics after immobilizing the molecules in an oriented fashion.

项目概要 动脉血栓形成发生在高液流条件下，并由以下物质的结合介导： 血小板通过其受体糖蛋白 Ib (GPIb) 转变成称为冯·维勒布兰德 (von Willebrand) 的血浆蛋白 通过 A1 域的因子 (VWF)。该过程通常会被下调以防止 自发粘连和血栓形成，但被高剪切应力激活。拟议的 这项工作将检验以下假设：GPIb 与 A1 的结合会被邻近的 A1 下调 VWF 内的域或侧翼区域，使得 VWF 显示域间自动抑制。它将 还检验了与高剪切应力相关的机械力激活的假设 通过将这些调节区域拉离 A1 结构域来缓解血小板结合 抑制。要测试的最终假设是增加自发性的条件 血小板与 VWF 的结合，包括 2B 型冯维勒布兰德病，是通过破坏 正常的域间自动抑制。 VWF 内的监管领域和要素将是 通过克隆 VWF 的不同区域并确定两者的粘附行为来识别 可溶性分子和表面固定分子。机械力和剪切应力的影响 该规定将在之后使用原子力显微镜和微流体来确定 以定向方式固定分子。