Molecular Basis for Platelet Dense Granule Biogenesis

血小板致密颗粒生物发生的分子基础

• 批准号: 6368517
• 负责人: ESTEBAN C DELL'ANGELICA
• 金额: $ 29.44万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-11 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6368517
• 关键词: biosynthesis; cell component structure /function; granule; laboratory mouse; membrane activity; membrane biogenesis; membrane proteins; platelets; protein protein interaction; protein purification; tissue /cell culture

DESCRIPTION (APPLICANT'S ABSTRACT): The long-term goal of this research is to understand at the molecular level the biogenesis of platelet dense granules. Deficiencies in these granules are associated with congenital and acquired disorders that manifest with prolonged bleeding. Understanding how platelet dense granules are formed, and which elements are involved in this process, may thus lead to the development of appropriate treatments for these disorders. In addition, the molecular machinery that generates platelet dense granules is a potential target for the development of drugs that could help reduce the risk of myocardial infarction and stroke. Although the mechanism of platelet dense granule biogenesis remains largely unknown, recent genetic studies have revealed that adaptor protein 3 (AP-3), a protein complex conserved among all eukaryotes, plays a critical role in this process. This finding has opened new avenues for the identification of additional elements involved. This application proposes to identify human proteins that interact physically with AP-3 and test the hypothesis that they are components of the molecular machinery that generates platelet dense granules. AP-3-interacting proteins will be isolated and characterized by biochemical methods, and their putative role in AP-3-dependent events will be assessed using a recently developed assay that allows detection of AP-3 function defects in various human cell lines. Subsequently, the direct involvement of these AP-3-interacting proteins in platelet dense granule biogenesis will be tested by mutational analyses in mouse models of platelet dense granule deficiency or, alternatively, by targeted disruption of the encoding gene in mice.

描述（申请人的摘要）：这项研究的长期目标是 在分子水平上了解血小板致密颗粒的生物发生。 这些颗粒的缺乏与先天性和后天性有关 表现为长期出血的疾病。了解血小板如何 形成致密颗粒，该过程涉及哪些元素，可能 从而导致开发出针对这些疾病的适当治疗方法。在 此外，产生血小板致密颗粒的分子机制是 开发有助于降低风险的药物的潜在目标 心肌梗塞和中风。 尽管血小板致密颗粒生物发生的机制在很大程度上仍然存在 未知，最近的遗传学研究表明，接头蛋白 3 (AP-3) 所有真核生物中保守的蛋白质复合物，在此过程中发挥着关键作用 过程。这一发现开辟了新的途径来识别 涉及的其他元素。该应用程序旨在识别人类 与 AP-3 发生物理相互作用的蛋白质并检验它们的假设 是产生血小板密集的分子机器的组成部分 颗粒。 AP-3 相互作用蛋白将被分离并表征为 生化方法，以及它们在 AP-3 依赖性事件中的假定作用将是 使用最近开发的可检测 AP-3 的测定法进行评估 各种人类细胞系的功能缺陷。随后，直接 这些 AP-3 相互作用蛋白参与血小板致密颗粒 生物发生将通过血小板小鼠模型的突变分析进行测试 致密颗粒缺陷，或者通过有针对性地破坏 小鼠的编码基因。