Investigation of Platelet G-Protein Coupled Receptors

血小板 G 蛋白偶联受体的研究

基本信息

批准号：
9097151
负责人：
Fadi T Khasawneh
金额：
$ 42.07万
依托单位：
WESTERN UNIVERSITY OF HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-23 至 2016-10-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9097151
关键词：
ADP Receptors Address Agonist Amino Acid Sequence Amino Acids Antibodies Binding Biological Biology Bleeding time procedure Blood Platelets Clinical Coupling Data Development Disease Dose Dot Immunoblotting Event Exhibits Foundations G-Protein-Coupled Receptors GTP-Binding Proteins Goals Health Hemostatic function Human IL2 gene Immunization In Vitro Investigation Knowledge Ligand Binding Ligand Binding Domain Maps Mediating Modeling Molecular Molecular Models Mus Nature Pathogenesis Pathway interactions Peptides Platelet Activation Platelet aggregation Play Prevention Publishing Reagent Receptor Signaling Resources Role Serotonin Shapes Signal Transduction Specificity Structure Testing Therapeutic Agents Thrombocytopenia Thrombosis Thromboxane A2 Time Vaccinated base design extracellular in vivo interest mimetics molecular modeling mouse model new therapeutic target novel peptide based vaccine peptidomimetics protein aminoacid sequence purinoceptor P2Y1 receptor receptor coupling research study structural biology

项目摘要

DESCRIPTION (provided by applicant): While the involvement of signaling of the serotonin 5HT2A receptor (5HT2AR) and the ADP P2Y1 receptor in the pathogenesis of thrombotic diseases is well documented, there are no antagonists that are currently available for clinical use, and which target either of these pathways. This derives, in part, from lack of knowledge regarding receptor signaling and structure. The present application proposes experiments that address fundamental aspects of the structural biology and signaling of the platelet 5HT2A and P2Y1 receptors: 1. the biological significance of antagonism of 5HT2AR ligand-binding region (i.e., the second extracellular loop; EL2) in the prevention of thrombotic diseases. Our hypothesis is that EL2 of 5HT2AR plays an important role in platelet activation. To address this hypothesis, our experiments will evaluate the ability of a hypothesis is that EL2 of 5HT2AR plays an important role in platelet activation. enhanced, agonist-induced platelet activation. Subsequent studies we will investigate the effects of EL2Ab and an EL2 peptide-based "vaccine" on bleeding time, and thrombosis development. 2. The G-protein coupling domains of P2Y1 receptor. Our hypothesis is that the intracellular domains of P2Y1 contain separate regions that confine coupling to a specific G-protein. In this regard, one of our interesting resuls is that a peptide mimicking the N-terminus of the second intracellular loop (IL2) of P2Y1 (abbreviated Myr-N-IL2pep) blocked ADP-induced aggregation. This finding suggests that the N-IL2 domain of P2Y1 participates in receptor coupling to Gq. Similarly, the role of other IL regions in G-protein coupling will be determined by examining the effects of their corresponding peptides on ADP-triggered platelet activation. Also, the effects of any biologically-active IL peptides on bleeding time and thrombosis development will be investigated. Collectively, results obtained from these studies will provide fundamental information concerning 5HT2AR and P2Y1 receptor biology and structure, and may define new therapeutic targets and/or aid molecular modeling study predictions for organic derivatives/agents for treating thrombotic disease states.

描述（由应用提供）：虽然血清素5HT2A受体（5HT2AR）和ADP P2Y1受体在血栓性疾病的发病机理中的信号传导涉及，但目前没有任何拮抗剂可用于临床使用，哪些针对这些途径。这部分源于缺乏有关受体信号传导和结构的知识。目前的应用建议实验，涉及血小板5HT2A和P2Y1受体的结构生物学的基本方面以及信号：1。5HT2AR配体结合区域（即第二个细胞外循环; EL2）的拮抗作用生物学性在预防螺栓脉络膜的预防中。我们的假设是5HT2AR的EL2在血小板激活中起重要作用。为了解决这一假设，我们的实验将评估假设的能力是5HT2AR的EL2在血小板激活中起重要作用。增强的，激动剂诱导的血小板激活。随后的研究，我们将研究EL2AB和基于EL2肽的“疫苗”对出血时间和血栓形成发育的影响。 2。P2Y1受体的G蛋白偶联结构域。我们的假设是，P2Y1的细胞内结构域包含局限于特定G蛋白的单独区域。在这方面，我们有趣的结果之一是，模仿P2Y1（缩写的MYR-N-IL2PEP）的第二个细胞内环（IL2）的N端阻断了ADP诱导的聚集。这一发现表明，P2Y1的N-IL2结构域参与受体耦合与GQ。同样，其他IL区域在G蛋白偶联中的作用将通过检查其相应的辣椒对ADP触发的血小板激活的影响来确定。同样，将研究任何生物活性IL对出血时间和血栓形成发展的影响。总的来说，从这些研究中获得的结果将提供有关5HT2AR和P2Y1受体生物学和结构的基本信息，并可能定义了新的治疗靶标和/或有助于分子建模研究预测有机衍生物/药物用于治疗血栓性疾病状态的预测。