Molecular mechanisms of coat assembly and regulation in membrane trafficking pathways

膜运输途径中外壳组装和调节的分子机制

• 批准号: 10405921
• 负责人: Lauren Parker Jackson
• 金额: $ 45.59万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10405921
• 关键词: Affinity; Alzheimer' s Disease; Animal Model; Binding; Biochemical; Capsid Proteins; Cell Line; Cellular Membrane; Cellular biology; Clathrin; Coat Protein Complex I; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Data; Defect; Destinations; Ensure; Event; Family; Goals; Health; Human; Integral Membrane Protein; Link; Lipids; Malignant Neoplasms; Membrane; Methods; Molecular; Molecular Structure; Movement; Parkinson Disease; Pathway interactions; Phenotype; Physiological Processes; Proteins; Regulation; SNAP receptor; Spastic Paraplegia; Structural Models; Testing; Time; Tubular formation; Vesicle; Work; X-Ray Crystallography; biophysical techniques; design; experimental study; genetic regulatory protein; human disease; insight; nervous system disorder; polypeptide; protein complex; protein protein interaction; protein transport; tool; trafficking

Project Summary The timely delivery of membrane-bound vesicles and tubules bearing transmembrane protein and lipid cargoes to discrete cellular membranes is fundamental to cell biology and human health. Many proteins associated with trafficking pathways are linked to serious and crippling human diseases, especially neurological diseases and disorders. Although many trafficking proteins and some pathways are well characterized, we still do not understand other trafficking pathways that we infer must exist between membranes. This constitutes an enormous gap in our understanding of fundamental cell biology. Our goal is to elucidate the molecular structures and functions of important coat protein complexes that initiate trafficking pathways by forming coats around vesicles or tubules. Coat proteins recognize and package relevant cargoes, and they promote efficient assembly of additional required protein components, like accessory proteins and SNAREs. Clathrin coats have long served as an important paradigm, but increasing evidence demonstrates how other coats use distinct mechanisms. We investigate the retromer and Assembly Polypeptide (AP) family of coat complexes (COPI, AP-4, AP-5) by using a variety of tools to ascertain molecular mechanisms of coat assembly and regulation. Biochemical approaches allow us to identify and test new interactions in coat complexes, including how accessory and regulatory proteins drive function. Integrated structural methods including X-ray crystallography, cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) provide detailed evidence for how coats interact with key partners and allow us to generate specific hypotheses to test function. Biophysical techniques enable us to quantify binding affinities and to probe interfaces identified in structural models. With collaborators, we use molecular data to design experiments in cultured cell lines or in model organisms to explore how protein-protein interactions drive phenotypes at the cellular or organismal levels. Ultimately, we hope to gain a molecular understanding of how coats assemble at distinct membranes to drive different trafficking pathways. We anticipate this work will reveal new mechanisms of coat assembly and regulation and will provide fundamental insights into the protein networks that underlie key cellular events on membranes.

项目摘要 携带跨膜蛋白和脂质货物的膜结合囊泡和小管的及时递送 分离细胞膜是细胞生物学和人类健康的基础。许多蛋白质与 贩运途径与严重和致残的人类疾病，特别是神经系统疾病有关， 紊乱虽然许多运输蛋白和一些途径的特点，我们仍然没有 了解我们推断必须存在于膜之间的其他运输途径。这构成了 我们对基本细胞生物学的理解存在巨大差距。我们的目标是阐明 通过形成外壳启动运输途径的重要外壳蛋白复合物的结构和功能 围绕着小泡或小管。外壳蛋白识别并包装相关货物，它们促进高效的运输。 组装额外所需的蛋白质组分，如辅助蛋白和SNARE。网格蛋白涂层具有 长期以来一直是一个重要的范例，但越来越多的证据表明，其他外套如何使用不同的 机制等我们研究了逆转录聚合物和组装多肽（AP）家族的外壳复合物（COPI， AP-4，AP-5）通过使用各种工具来确定外套组装和调节的分子机制。 生物化学方法使我们能够识别和测试新的相互作用的外套复合物，包括如何 辅助和调节蛋白驱动功能。综合结构方法，包括X射线晶体学， 冷冻电子显微镜（cryo-EM）和冷冻电子断层扫描（cryo-ET）提供了 涂层如何与关键伙伴互动，并允许我们生成特定的假设来测试功能。生物物理 技术使我们能够量化结合亲和力和探测结构模型中识别的界面。与 合作者，我们使用分子数据来设计培养细胞系或模型生物的实验， 探索蛋白质-蛋白质相互作用如何在细胞或生物体水平上驱动表型。最终我们 希望从分子水平上了解外套是如何在不同的细胞膜上组装， 贩运途径。我们预计这项工作将揭示被毛组装和调节的新机制， 将提供对膜上关键细胞事件的蛋白质网络的基本见解。