MOLECULAR BASIS FOR PLATELET FUNCTION IN HEMOSTASIS

该项目由止血中的血小板功能分子基础罗伯特·W·科尔曼指导，将在坦普尔大学医学院的索尔·雪莉血栓研究中心进行。该中心是医学院内正式成立的研究机构，致力于促进我们对动、静脉血栓形成和出血性疾病的病因/发病机制、诊断和治疗的了解。为了实现这一目标，该项目的研究人员将研究血小板在止血和血栓形成中的激活机制。本提案中的每个项目都有可能表明或为开发适当的抑制药物或治疗方法提供基础，以防止血小板激活、聚集和凝血活性。和第二信使，特别是环核苷酸和血小板凝血活性。项目1，cAMP磷酸二酯酶3A的活性部位氨基酸(R.W.Colman)，集中在血小板中主要cAMP磷酸二酯酶活性部位的关键氨基酸的划定上。对3A型PDE的催化和金属结合至关重要的氨基酸将通过亲和标记和定点突变来鉴定。项目2，血小板因子XI(P.N.Walsh)，将从结构上表征血小板因子XI，并研究激动剂在血小板中暴露于激活因子XI的机制。我们将研究血小板因子XI的激活方式、细胞定位和功能特性。项目3，人类血小板中的信号转导缺陷(A.K.RAO)，将通过对一组质量缺陷患者的生化和分子生物学解剖，寻求对血小板信号转导机制的新见解，这些缺陷集中在PLC-B1a和Gaq的异常上。项目4，血小板ADP受体(S.P.ukapuli)，将阐明3种ADP受体亚型在血小板纤维蛋白原受体激活中的作用。核心A，行政(R.W.科尔曼)，将为所有项目提供财政监督和文书支持。核心B，细胞培养(S.P.库纳普利)将促进和支持所有项目，从而有效地加快研究。核心C，流式细胞仪(J.K.DeRiel)，将为所有项目提供荧光激活的细胞分析和共聚焦服务。所有这些调查人员都有与不止一名其他项目负责人进行合作研究的历史，并将与其他项目负责人协商和共享智力和物质资源。从这些研究中获得的知识将为加强止血和预防血栓形成提供治疗方法。

This Program Project, under the direction of Robert W. Coleman, Molecular Basis for Platelet Function in Hemostasis, will exist within the Sol Sherry Thrombosis Research Center of Temple University School of Medicine. The Center is a formally established research institute within the School of Medicine dedicated to advancing our understanding of the etiology/pathogenesis, diagnosis and treatment of arterial and venous thrombosis and hemorrhagic diseases. To fulfill this goal, investigators in this project will examine the mechanisms of activation of platelets in hemostasis and thrombogenesis. Each project in the present proposal has the potential to indicate or provide the foundation for development of an appropriate inhibitory drug or therapeutic approach to prevent platelet activation, aggregation, and coagulant activity. and second messengers in particular, cyclic nucleotides and platelet coagulant activities. Project 1, Active Site Amino Acids of cAMP Phosphodiesterase 3A (R.W. Colman), concentrates on the delineation on the critical amino acids in active sites of the major cAMP phosphodiesterase in platelet. The amino acids critical for catalysis and metal binding of type 3A PDE will be identified by affinity labeling and site-directed mutagenesis. Project 2, Platelet Factor XI (P.N. Walsh), will structurally characterize platelet factor XI and study the mechanism of agonist exposure of actor XI in platelets. The mode of activation, cellular localization, and functional characteristics of platelet factor XI will be investigated. Project 3, Signal Transduction Defects in Human Platelets (A.K. Rao), will seek new insights into platelet signal transduction mechanisms by the biochemical and molecular biological dissection of the defects of a selected group of patients with qualitative defects focusing on abnormalities in PLC-b1a and Gaq. Project 4, Platelet ADP Receptors (S.P. Kunapuli), will elucidate the role of 3 ADP receptor subtypes in platelet fibrinogen receptor activation. Core A, Administration (R.W. Colman), will provide fiscal oversight and clerical support for all projects. Core B, Cell Culture (S.P. Kunapuli), will facilitate and support all of the projects and thus efficiently expedite the research. Core C, Flow Cytometry (J.K. deRiel), will provide fluorescent-activated cell analysis and confocal services to all projects. All of these investigators have a history of collaborative research with more than one of the other project leaders and will consult and share intellectual and physical resources with the other project leaders. The knowledge gained from these studies will provide therapeutic approaches to enhancing hemostasis and preventing thrombosis.