Proteins of Coagulation Pathways

凝血途径的蛋白质

• 批准号: 7930567
• 负责人: JOHN H GRIFFIN
• 金额: $ 58.22万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7930567
• 关键词: 3-Dimensional; Abbreviations; Affect; Anticoagulants; Arteries; Basic Science; Binding; Binding Proteins; Biological Assay; Biological Markers; Blood; Blood Clot; Blood Coagulation Factor; Blood coagulation; Carnitine; Chimera organism; Cholesterol Esters; Choline; Clinical; Clinical Research; Coagulation Process; Complex; Computer Simulation; Crystallography; Data; Diagnosis; Disease; Dyslipidemias; Elements; Energy Transfer; Factor Va; Factor X; Factor XII; Factor Xa; Fatty acid glycerol esters; Freezing; Functional disorder; Funding; Generations; Glucosylceramides; Glutamic Acid; Glycerol; Goals; Knowledge; Label; Leg; Life; Ligand Binding Domain; Link; Lipid Binding; Lipids; Lipoproteins; Lung; Lysophospholipids; Maps; Membrane; Minor; Modeling; Molecular; Mutation; Myocardial Infarction; Pathway interactions; Patients; Phospholipid Transfer Proteins; Phospholipids; Plasma; Plasma Proteins; Property; Protein Binding; Proteins; Prothrombin; Recombinants; Registries; Regulation; Risk; Sampling; Serine; Serum amyloid A protein; Site; Solutions; Sphingolipids; Sphingosine; Study Subject; Sulfoglycosphingolipids; Surface; Surface Plasmon Resonance; Testing; Thrombin; Thrombophilia; Thromboplastin; Thrombosis; Transfer Factor; Translating; Translational Research; Triglycerides; Variant; Veins; Venous; Venous Thrombosis; base; bench to bedside; clinically relevant; improved; innovation; insight; lipid transfer protein; lysophosphatidic acid; meizothrombin; member; novel; plasma phospholipid transfer protein; progesterone 11-hemisuccinate-(2-iodohistamine); prothrombinase complex; sialogangliosides

This revised renewal two year ARRA-funded project extends the PI's basic and clinical research on regulation of blood coagulation and on translational clinical research of venous thrombosis. Thrombosis is strongly linked to an imbalance of anticoagulant and procoagulant mechanisms. Three of four specific aims involve basic studies of novel plasma molecules that regulate clotting while the last aim involves translational research to identify novel biomarkers for thrombosis. For aims 1 and 2, we hypothesize that thrombin generation can be influenced by minor abundance single chain plasma lipids, so-called "soluble" lipids. Based on Surface Plasmon Resonance (SPR) binding studies and on clotting assays, we hypothesize that anticoagulant acyl carnitines, including palmitoyl carnitine, directly inhibit coagulation factor Xa by binding to Xa. In preliminary studies, palmitoyl carnitine binds to Gla-domainless-factor Xa and inhibits its activity. These preliminary data and the proposed experimentation thus relate to a novel paradigm for the effects of plasma lipids on coagulation pathways. Recombinant factor X/IX chimeras and variant factor X molecules from various species will be used to identify putative lipid binding domains on factor Xa. For aim 2, we hypothesize that plasma phospholipid transfer protein (PLTP) can directly influence plasma coagulability and thrombin generation by novel direct interactions with clotting factors. SPR preliminary data show that PLTP binds to specific clotting factors. For aim 3, we will determine some of the three dimensional structural properties of the prothrombinase complex comprising factors Xa, Va and prothrombin on phospholipid membranes. First, we will introduce Cys mutations and prepare fluorescently labeled factor Va. Then we will use Forster Resonance Energy Transfer (FRET) to generate a set of multiple point- to-point and point-to-plane distances that can be used to generate and then interpret FRET-derived distances for the prothrombinase complex. For aim 4, based on the hypothesis that imbalances of plasma anticoagulant minor abundance lipids are linked to thrombosis risk, we will use already available frozen plasma samples from thrombosis patients and matched controls to determine if certain targeted plasma lipids or two lipid binding plasma proteins (PLTP or serum amyloid A) are biomarkers for thrombosis. This project will increase insights into the pathophysiology of thrombosis and may improve diagnosis and treatment of thrombosis.

这项修订后的两年ARRA资助项目扩展了PI的基本和临床 关于血液凝血调节的研究以及关于转化的临床研究 静脉血栓形成。血栓形成与抗凝剂的失衡密切相关， Procagulant机制。四个特定目标中的三个涉及新颖的基础研究 调节凝结的血浆分子，而最后一个目标涉及转化研究 鉴定新型生物标志物用于血栓形成。对于目标1和2，我们假设 凝血酶产生可能受到次要丰度单链血浆脂质的影响， 所谓的“可溶”脂质。基于表面等离子体共振（SPR）结合研究 在凝血测定中，我们假设 棕榈酰肉碱，通过与Xa结合直接抑制凝血因子Xa。在初步 研究，棕榈酰肉碱与GLA无域因子Xa结合并抑制其活性。 这些初步数据和提出的实验因此与新型有关 血浆脂质对凝血途径的影响的范例。重组因子 X/IX嵌合体和变体因子X分子将用于 识别因子XA上推定的脂质结合域。对于目标2，我们假设 血浆磷脂转移蛋白（PLTP）可以直接影响血浆凝血性 通过与凝血因子进行新型直接相互作用的凝血酶生成。 spr 初步数据表明，PLTP与特定的凝血因子结合。对于目标3，我们将 确定凝血酶酶的三维结构特性中的某些 复杂的磷脂膜上包含Xa，VA和凝血酶原的因子。第一的， 我们将引入CYS突变并准备荧光标记的因子VA。然后我们 将使用Forster共振能量传递（FRET）生成一组多点 - 可用于生成然后解释的点对点距离 凝结蛋白酶复合物的fret衍生距离。对于AIM 4，基于 假设血浆抗凝剂少脂质的失衡与 血栓形成风险，我们将使用血栓形成的已经可用的冷冻血浆样品 患者和匹配的对照，以确定某些靶向血浆脂质还是两种脂质 结合血浆蛋白（PLTP或血清淀粉样蛋白A）是血栓形成的生物标志物。这 项目将增加对血栓形成的病理生理学的见解，并可能改善 血栓形成的诊断和治疗。