REGULATION OF PLATELET FUNCTION BY GPIIB-IIIA

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。研究将使用健康志愿者。将抽取血液并分离血小板。评估钙蛋白酶、血影蛋白和血小板生成素作用的实验将涉及抑制血小板内的特定途径和相互作用并评估功能后果。将用特定的药理学抑制剂和通过引入棕榈酰化肽来抑制途径。后一种肽将被设计为消除感兴趣的相互作用，例如血影蛋白SH 3结构域与鉴定的结合配偶体之间的相互作用，血影蛋白与整联蛋白之间的相互作用，或钙蛋白酶与鉴定的结合配偶体之间的相互作用。通过测量整联蛋白aIIbb 3被激活的能力、血小板粘附的能力或血小板扩散的能力来评估消融特异性信号分子或相互作用的后果。将通过免疫荧光研究来评估抑制的详细效果，所述免疫荧光研究鉴定了铺展细胞中整联蛋白、肌动蛋白丝、血影蛋白、钙蛋白酶和肌动蛋白的再分布。在其他研究中，血小板将用于特定信号分子（如钙蛋白酶、Rac、cdc 42）的测定。将从血小板中分离纯化的蛋白质，用于体外结合试验、产生抗体和鉴定钙蛋白酶裂解产物的体外试验。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Healthy volunteers will be used for the studies. Blood will be drawn and platelets isolated. Experiments to assess the role of calpain, spectrin, and skelemin will involve inhibiting specific pathways and interactions within platelets and assessing the functional conseqences. Pathways will be inhibited with specific pharmacological inhibitors and by the introduction of palmitylated peptides. The latter peptides will be designed to ablate interactions of interest, such as those between spectrin SH3 domain and identified binding partners, that between skelemin and integrin, or that between calpain and identified binding partners. Consequence of ablating specific signaling molecules or interactions will be assessed by meausuring the ability of the integrin aIIbb3 to be activated, the ability of platelets to adhere or the ability of platelets to spread. Detailed effects of inhibition will be assessed by immnofluorescent studies identifying the redistribution of integrin, actin filaments, spectrin, calpain and skelemin in spreading cells. In other studies, platelets will be used for assays of specific signaling molecules such as calpain, Rac, cdc42. Isolation of purified proteins from platelets will be performed for in vitro binding assays, raising antibodies, and for in vitro assays identifying calpain cleavage products.