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（由申请人提供）：尽管血清素 5HT2A 受体 (5HT2AR) 和 ADP P2Y1 受体的信号传导参与血栓性疾病的发病机制已得到充分证明，但目前尚无可用于临床的拮抗剂，并且其靶向这部分源于缺乏关于受体信号传导和结构的知识。本申请提出了解决血小板的结构生物学和信号传导的基本方面的实验。 5HT2A和P2Y1受体：1.拮抗5HT2AR配体结合区（即第二细胞外环；EL2）在预防血栓性疾病中的生物学意义我们的假设是5HT2AR的EL2在血小板活化中发挥重要作用。为了解决这个假设，我们的实验将评估一个假设的能力，即 5HT2AR 的 EL2 在血小板活化增强中发挥重要作用。后续研究我们将研究 EL2Ab 和基于 EL2 肽的“疫苗”对出血时间和血栓形成的影响。我们的假设是细胞内的 G 蛋白偶联结构域。 P2Y1 的结构域包含限制与特定 G 蛋白偶联的单独区域。在这方面，我们有趣的结果之一是模拟第二个细胞内环的 N 末端的肽。 P2Y1 的 (IL2)（缩写为 Myr-N-IL2pep）可阻断 ADP 诱导的聚集，这一发现表明 P2Y1 的 N-IL2 结构域参与与 Gq 的受体偶联，类似地，其他 IL 区域在 G 蛋白偶联中也发挥着作用。将通过检查其相应肽对 ADP 触发的血小板活化的影响来确定。此外，任何生物活性 IL 肽对出血时间和血栓形成的影响也将被确定。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，并可能定义新的治疗靶点和/或帮助分子模型研究预测用于治疗血栓性疾病状态的有机衍生物/药剂。