CRYSTALLOGRAPHIC ANALYSIS OF SYNAPTOTAGMIN C2A-C2B

突触结合蛋白 C2A-C2B 的晶体分析

• 批准号: 7954337
• 负责人: ROGER BRYAN SUTTON
• 金额: $ 0.35万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954337
• 关键词: Area; Binding; Biochemical; C2 Domain; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Funding; Grant; Human; Institution; Mediating; Methods; Molecular Conformation; Neurons; Neurotransmitters; Phospholipids; Protein Binding; Research; Research Personnel; Resolution; Resources; SNAP receptor; Source; Structure; United States National Institutes of Health; Vesicle; complement C2a; complement C2b; design; presynaptic; structural biology; synaptotagmin; synaptotagmin I; synchrotron radiation

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. Synaptotagmin 1 mediates the Ca+2-dependent fusion of neurotransmitter-filled vesicles with the presynaptic terminus in neurons. While there is considerable structural and biochemical information on the synaptotagmin molecule, its precise function is still unclear; however, most atomic resolution structures conclude that the two tandem C2 domains of synaptotagmin were autonomous agents attached by a small linker. Due to our unique purification and crystallization method, we have captured human synaptotagmin 1 C2A-C2B in a conformation that shows contacts between domains as they exist prior to Ca+2 priming. The crystals we have analyzed at SSRL to this point reveal that areas of C2A involved in Ca+2 binding, phospholipid association and SNARE protein binding are distorted and probably inactivated through intra-molecular interactions with C2B. These interactions form a trigger mechanism designed to respond to changing Ca+2 concentrations. These conclusions will be essential to our understanding of fundamental neuronal function.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 Synaptotagmin 1 介导充满神经递质的囊泡与神经元突触前末端的 Ca+2 依赖性融合。 虽然关于突触结合蛋白分子有大量的结构和生化信息，但其确切功能仍不清楚。然而，大多数原子分辨率结构得出的结论是，突触结合蛋白的两个串联 C2 结构域是由一个小接头连接的自主代理。 由于我们独特的纯化和结晶方法，我们捕获了人突触结合蛋白 1 C2A-C2B，其构象显示结构域之间在 Ca+2 引发之前存在的接触。 到目前为止，我们在 SSRL 分析的晶体表明，参与 Ca+2 结合、磷脂缔合和 SNARE 蛋白结合的 C2A 区域被扭曲，并且可能通过与 C2B 的分子内相互作用而失活。 这些相互作用形成了一种触发机制，旨在响应 Ca+2 浓度的变化。 这些结论对于我们理解基本神经元功能至关重要。