STRUCTURE/FUNCTION OF THE SNARE COMPLEX

圈套复合体的结构/功能

• 批准号: 6477236
• 负责人: Phyllis I Hanson
• 金额: $ 22.76万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-03 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6477236
• 关键词: SDS polyacrylamide gel electrophoresis; Saccharomyces cerevisiae; electron microscopy; eukaryote; exocytosis; freeze etching; immunoprecipitation; intracellular membranes; intracellular transport; laboratory rat; membrane fusion; membrane permeability; membrane proteins; neuronal transport; physical model; protein binding; protein protein interaction; protein sequence; protein structure function; syntaxin

The long-term goal of this project is to understand the molecular reactions that appear to be responsible for neuronal exocytosis, and more generally for membrane trafficking throughout eukaryotic cells. A number of proteins that participate in exocytosis have been identified, including the synaptic vesicle protein synaptobrevin and the plasma membrane proteins syntaxin and SNAP-25. These three proteins are referred to collectively as SNAREs (soluble NSF-attachment protein receptors), and are representative of families of related proteins that function at different membrane trafficking steps throughout the cell. The three SNAREs bind tightly to each other to form a SNARE complex that is thought to interconnect membranes destined to fuse with each other. SNARE complexes are stable unless dissociated by the ATPase NSF (N-ethylmaleimide sensitive fusion protein), and cycle assembly and disassembly of SNARE complexes is correlated with membrane fusion in vivo. However, it remains unclear exactly how these reactions are linked to membrane fusion. In the proposed experiments, we will determine how the SNARE complex participates in membrane fusion by generating soluble SNARE complexes, delineating their structure, and characterizing their interactions with other critical complexes in solution will lead up to predictions about how such complexes affect membranes in vivo. We ill test these predications using 'model neuronal membranes' produced by heterologously expressing the synaptic SNAREs on secretory vesicles in the yeast Saccharomyces cerevisiae, isolating these vesicles, and studying their interactions using biochemical and ultrastructural approaches. With this information we will be better able to understand how intracellular membrane fusion, and in particular neurosecretion, can be modulated or abolished and how such alterations contribute to human disease and could be exploited to therapeutic advantage.

该项目的长期目标是了解分子 反应似乎是负责神经元胞吐，以及更多 通常用于整个真核细胞的膜运输。一些 参与胞吐作用的蛋白质已经被鉴定，包括 突触囊泡蛋白synaptobrevin和质膜 蛋白质syntaxin和SNAP-25。这三种蛋白质被称为 统称为SNARE（可溶性NSF附着蛋白受体）， 代表相关蛋白质家族的蛋白质， 膜运输步骤贯穿整个细胞。这三个陷阱 彼此紧密相连，形成一个被认为是 互连膜，这些膜注定要彼此融合。SNARE复合物 除非被ATP酶NSF（N-乙基马来酰亚胺）解离，否则是稳定的 敏感的融合蛋白），以及SNARE的循环组装和拆卸 复合物与体内膜融合相关。但委员会仍 目前还不清楚这些反应是如何与膜融合联系在一起的。在 建议的实验，我们将确定如何陷阱复杂参与 在膜融合中通过产生可溶性SNARE复合物， 它们的结构，并描述它们与其他关键 溶液中的复合物将导致预测这些复合物如何 影响体内膜。我们将使用“模型”来检验这些预测 通过异源表达突触蛋白而产生的神经元膜 酿酒酵母分泌囊泡上的SNARE， 分离这些囊泡，并研究它们之间的相互作用， 生化和超微结构方法。有了这些信息，我们将 能够更好地理解细胞内膜融合， 特别是神经分泌，可以调节或消除，以及如何这样做， 改变有助于人类疾病，并可以利用 治疗优势