STRUCTURE/FUNCTION OF THE SNARE COMPLEX

圈套复合体的结构/功能

• 批准号: 6330558
• 负责人: Phyllis I Hanson
• 金额: $ 22.1万
• 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 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6330558
• 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.

这个项目的长期目标是了解分子 似乎与神经元胞吐有关的反应，以及更多 通常用于真核细胞的膜运输。一个数字 已经确定了参与胞吐作用的蛋白质，包括 突触囊泡蛋白Synaptobrevin与质膜 蛋白质合成素和SNAP-25。这三种蛋白质指的是 统称为SNARS(可溶性NSF附着蛋白受体)，并且是 具有不同功能的相关蛋白质家族的代表 膜运输遍及整个细胞。三个陷阱捆绑在一起 彼此紧紧地靠在一起，形成了一种被认为是 相互连接的膜注定要彼此融合。圈套复合体 除非被ATPase NSF(N-乙基马来酰亚胺)解离，否则是稳定的 敏感融合蛋白)，以及SNARE的循环组装和拆解 复合体与体内膜融合相关。然而，它仍然 目前尚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，我们将确定陷阱复合体是如何参与的 在膜融合中通过产生可溶的SNAR络合物，描绘 它们的结构，并表征它们与其他关键 溶液中的络合物将导致对这些络合物如何 影响体内的细胞膜。我们将使用‘模型’来测试这些预测 异源表达突触所产生的神经细胞膜 诱捕酵母中的分泌囊泡， 分离这些囊泡，并研究它们的相互作用 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是如何进行的 特定的神经分泌，可以被调节或取消，以及如何调节或取消 基因改变会导致人类疾病，并可能被利用来 治疗优势。