Dynamics of Endomembrane Docking and Fusion

内膜对接和融合的动力学

基本信息

批准号：
9893719
负责人：
Alexey Jarrell Merz
金额：
$ 34万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9893719
关键词：
3-Dimensional Architecture Binding Biochemical Biochemical Genetics Biological Models Biophysics Bypass Cells Charcot-Marie-Tooth Disease Communicable Diseases Complex Coupled Cryoelectron Microscopy Degenerative Disorder Development Diabetes Mellitus Docking Ebola virus Endocrine Eukaryotic Cell Family Frankfurt-Marburg Syndrome Virus Functional disorder Genetic Goals Golgi Apparatus Homeostasis Human Immune signaling Immunity In Vitro Lipids Mass Spectrum Analysis Mediating Membrane Membrane Fusion Membrane Protein Traffic Methods Modeling Molecular Molecular Chaperones Nerve Degeneration Neurodegenerative Disorders Neutropenia Organelles Phase Process Proteins Quantitative Genetics Reaction Regulation Reporting Research Personnel Resolution Roentgen Rays Role SNAP receptor Saccharomyces cerevisiae Saccharomycetales Somatic Cell Genetics Structure System Techniques Testing Variant Virus Diseases Work Yeasts alpha-SNAP biophysical techniques cofactor comparative crosslink detection of nutrient experimental study genetic analysis in vitro Assay in vivo innovation interest mutation screening neurotransmission novel premature professor protein activation protein function protein transport proteoliposomes public health relevance reconstitution stoichiometry syntaxin binding protein 1 trafficking vesicle transport

项目摘要

Project Summary Membrane traffic is among the most ancient innovations of eukaryotic cells. It is central to neurotransmission, immune signaling, and normal development, and disrupted in a wide array of infectious and degenerative diseases. The fusion of transport vesicles with target membranes is central to many trafficking processes and employs a core and conserved machinery: three or four SNARE proteins, the disassembly chaperones Sec17 (α-SNAP) and Sec18 (NSF), and proteins of the SM (Sec1/Munc18) family. The mechanism of SM protein function is not understood, though several SM proteins are associated with human infectious disease, neurodegeneration, neutropenia, and diabetes. The overall goal of this Project is to scrutinize the emerging hypothesis that the biochemical basis of SM function entails not only SM–SNARE interactions but an elegant network of physical and functional interactions among SNAREs, SMs, Sec17, and Sec18, operating in tightly coupled assembly and disassembly reactions. Much of the recent work on SM proteins and Sec17 has been done in vitro. This Project combines powerful in vitro biochemical approaches with state-of-the-art in vivo structure–function analyses that are currently feasible only in Saccharomyces cerevisiae. In Aim 1, the mechanisms of SM protein activation and activity are assessed in vivo and in vitro. In Aim 2, Sec17 interactions with SNAREs, Sec18, and specific SMs are assessed. In Aim 3, biophysical techniques are applied to elucidate the architecture of SM assembly with SNAREs and with Sec17. These studies will test general and deep hypotheses about the conserved mechanisms of SNARE-mediated membrane fusion, with particular emphasis on comparisons between two different transport steps, and between in vitro results and stringent and quantitative in vivo structure– function analyses.

项目摘要膜交通是真核细胞中最古老的创新之一。这是核心神经传递，免疫信号和正常发育，并在广泛的传染性和退化性疾病。运输蔬菜与目标膜的融合至中心许多贩运过程和员工是核心和构成机械的：三到四个圈套蛋白，拆卸伴侣SEC17（α-SNAP）和SEC18（NSF），以及SM的蛋白质（SEC1/MUNC18）家庭。尽管有几种SM蛋白是与人类传染病，神经退行性疾病，神经退行性，神经减少症和糖尿病有关。总体目标这个项目的是审查新兴的假设，即SM功能的生化基础不需要只有SM – Snare互动，但在实质性和功能相互作用之间的优雅网络贪食，SMS，SEC17和SEC18在紧密耦合的组件和拆卸反应中运行。大部分最近的SM蛋白和SEC17的工作是在体外完成的。该项目结合了强大的体外使用最新的体内结构 - 功能分析的生化方法目前可行仅在酿酒酵母中。在AIM 1中，SM蛋白激活和活性的机制是在体内和体外评估。在AIM 2中，SEC17与Snares，Sec18和特定SMS的相互作用是评估。在AIM 3中，采用生物物理技术来阐明SM组装的结构贪食和Sec17。这些研究将检验有关保守的一般和深层假设圈圈介导的膜融合的机制，特别强调了两个之间的比较不同的运输步骤以及体外结果与体内结构严格且定量的范围之间功能分析。