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: 8389604
• 负责人: Wendy E Thomas
• 金额: $ 39.91万
• 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/8389604
• 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.

项目摘要 动脉血栓形成发生在高流体流动条件下，并由 通过其受体糖蛋白IB（GPIB）的血小板到称为Von Willebrand的血浆蛋白 因子（VWF）通过A1域。这个过程通常会下调以防止 自发粘附和血栓形成，但被高剪切应力激活。提议 工作将检验以下假设：GPIB与A1结合被邻近下调 VWF内部的域或侧翼区域，因此VWF显示域间自动抑制。会 还测试了与高剪切应力相关的机械力激活的假设 血小板结合通过将这些调节区域拉开从A1域中拉开以减轻自动 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Von Willebrand疾病，通过损害 正常的域间自动抑制。 VWF内的监管域和元素将是 通过克隆VWF的不同区域并确定两者的粘附行为来识别 可溶性和表面不明的分子。机械力和剪切应力对 该调节将使用原子力显微镜和微流体进行确定 以定向方式固定分子。