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: 8771444
• 负责人: Wendy E Thomas
• 金额: $ 41.29万
• 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 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8771444
• 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的条件下，并且通过结合 血小板通过其受体糖蛋白Ib（GPIb）转化为血浆蛋白，称为von Willebrand 因子（VWF）通过A1结构域。这一过程通常被下调，以防止 自发粘附和血栓形成，但被高剪切应力激活。拟议 这项工作将检验GPIb与A1的结合被邻近的蛋白质下调的假设。 结构域或VWF内的侧翼区域，使得VWF显示结构域间自抑制。它将 还检验了与高剪切应力相关的机械力激活 通过将这些调节区域从A1结构域拉离，以减轻血小板的自动结合， 抑制作用最后一个有待检验的假设是， 血小板与VWF的结合，包括2B型血管性血友病，是通过破坏 正常的域间自抑制。VWF中的监管领域和要素将 通过克隆VWF的不同区域并确定两者的粘附行为来鉴定 可溶的和表面固定的分子。机械力和剪切应力对 这种调节将使用原子力显微镜和微流体来确定， 以定向方式固定分子。

项目成果

Structural basis of type 2A von Willebrand disease investigated by molecular dynamics simulations and experiments.

• DOI: 10.1371/journal.pone.0045207
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Interlandi G;Ling M;Tu AY;Chung DW;Thomas WE
• 通讯作者: Thomas WE

A multiplexed magnetic tweezer with precision particle tracking and bi-directional force control.

• DOI: 10.1186/s13036-017-0091-2
• 发表时间: 2017
• 期刊: Journal of biological engineering
• 影响因子: 5.6
• 作者: Johnson KC;Clemmens E;Mahmoud H;Kirkpatrick R;Vizcarra JC;Thomas WE
• 通讯作者: Thomas WE

Differential surface activation of the A1 domain of von Willebrand factor.

• DOI: 10.1116/1.4943618
• 发表时间: 2016-06-11
• 期刊: Biointerphases
• 影响因子: 2.1
• 作者: Tronic EH;Yakovenko O;Weidner T;Baio JE;Penkala R;Castner DG;Thomas WE
• 通讯作者: Thomas WE

The catch bond mechanism between von Willebrand factor and platelet surface receptors investigated by molecular dynamics simulations.

• DOI: 10.1002/prot.22759
• 发表时间: 2010-08-15
• 期刊: Proteins
• 影响因子: 2.9
• 作者: Interlandi G;Thomas W
• 通讯作者: Thomas W