Structure and Function of a Platelet SNARE Complex

血小板 SNARE 复合体的结构和功能

• 批准号: 6538100
• 负责人: Steven J. Freedman
• 金额: $ 12.49万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6538100
• 关键词: SDS polyacrylamide gel electrophoresis; X ray crystallography; chimeric proteins; circular dichroism; exocytosis; granule; membrane fusion; membrane proteins; molecular cloning; platelet activation; platelets; protein biosynthesis; protein structure function; secretion; stoichiometry; synaptic vesicles

DESCRIPTION (provided by applicant) This proposal is designed to merge the candidate's background in biochemistry and clinical experience in Hemostasis and Thrombosis to produce an independent physician -scientist investigator. Platelets are critical for maintaining blood vessel integrity in response to injury, but they are also responsible for acute arterial occlusion in atherosclerotic diseases, including myocardial infarction, stroke, and peripheral vascular disease. During platelet activation, alpha granules are secreted and release important molecules for platelet function, including thrombus formation. The molecular mechanism of secretion is thought to involve interactions between alpha granule and plasma membrane SNARE proteins (N-ethylmaleimide sensitive fusion protein receptors) to form a membrane fusion complex. There are three long-term aims of this research proposal: 1. To determine the assembly, stability, and stoichiometry of the alpha granule SNARE complex; 2. To determine the membrane fusion function of the alpha granule SNARE complex; 3. To determine the high resolution X-ray crystal structure of the alpha granule SNARE complex. Secondary aims will be to determine essential SNARE sequence requirements for function, and to determine the specificity of platelet SNARE interactions compared to those in other secretory cells. The methods used to achieve these goals will include a variety of biophysical and structural techniques, including circular dichroism spectroscopy, sedimentation equilibrium analysis, fluorescence spectroscopy, and X-ray crystallography. The laboratory for this research has special expertise in studying related proteins in this way. These studies will be fundamentally important to our understanding of alpha granule secretion in platelets, and will contribute significantly to SNARE protein biology by providing insight into the specificity of membrane fusion.

描述（由申请人提供） 该建议旨在合并候选人的生物化学背景 以及止血和血栓形成的临床经验，产生独立 医师 - 科学家研究员。 血小板对于维护至关重要 响应伤害的血管完整性，但它们也负有责任 用于动脉粥样硬化疾病的急性动脉闭塞，包括心肌 梗塞，中风和周围血管疾病。 在血小板期间 激活，α颗粒分泌，并释放重要的分子 血小板功能，包括血栓形成。 分子机制 分泌被认为涉及α颗粒与等离子体之间的相互作用 膜snare蛋白（N-乙基甲酰胺敏感融合蛋白受体） 形成膜融合复合物。 这有三个长期目标 研究建议：1。确定组装，稳定性和化学计量学 阿尔法颗粒小军团综合体； 2。确定膜融合 Alpha颗粒军团复合物的功能； 3。确定高 分辨率X射线晶体结构的Alpha颗粒SNARE复合物。 次要目的是确定基本的圈套序列要求 功能，并确定血小板相互作用的特异性 与其他分泌细胞中的细胞相比。 用于实现这些的方法 目标将包括各种生物物理和结构技术， 包括圆形二色性光谱，沉积平衡分析， 荧光光谱和X射线晶体学。 实验室 研究在以这种方式研究相关蛋白质方面具有特殊的专业知识。 这些研究对我们对alpha的理解至关重要 血小板中的颗粒分泌物，将为军鼓做出重大贡献 通过洞悉膜融合的特异性，蛋白质生物学。