Mechanism of secretory cargo export from the TGN

TGN 秘密货物出口机制

• 批准号: 10389640
• 负责人: Julia von Blume
• 金额: $ 12.5万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-12 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10389640
• 关键词: Actins; Address; Autoimmune Diseases; Binding Proteins; Ca(2+)-Transporting ATPase; Cell Compartmentation; Cell secretion; Cell surface; Cells; Coupled; Cytoskeleton; Defect; Destinations; Fluorescence; Golgi Apparatus; Homeostasis; Human; Hydrolase; IGF Type 2 Receptor; In Vitro; Lead; Membrane; Microscopy; Modernization; Organelles; Pathway interactions; Process; Proteins; Research; Resolution; Sorting - Cell Movement; Techniques; Tissues; base; genome editing; innovation; insight; malignant neurologic neoplasms; membrane model; nervous system disorder; protein distribution; protein transport; reconstitution; secretory protein; trans-Golgi Network

Project Summary Sorting of soluble proteins for transport to intracellular compartments and secretion from cells is essential for cell and tissue homeostasis. A major regulator of intracellular protein distribution is the trans-Golgi network (TGN) that sorts secretory proteins into specific carriers to transport them to their final destination. The sorting of lysosomal hydrolases at the TGN by the mannose-6-phosphate receptor is well established. However, the sorting mechanism for secreted proteins remains poorly understood. The recent discovery of a Ca2+-based sorting of secretory cargo at the TGN is beginning to uncover the mechanism by which cells sort secretory cargoes from Golgi residents and cargoes destined to the other cellular compartments. This Ca2+ based sorting involves the cytoplasmic actin cytoskeleton, which through membrane-anchored Ca2+ ATPase SPCA1 and the luminal Ca2+ binding protein Cab45 sorts of a subset of secretory proteins at the TGN. The underlying mechanism regulating the interplay between these components, however, is not clear. We take an interdisciplinary and highly innovative approach to address this issue of fundamental importance. Our aims include in vitro reconstitution of these components in model membranes to identify the minimal machinery required for the sorting of secretory cargoes. Modern genome editing techniques in human cells coupled to fluorescence and super-resolution microscopy will provide new insights into the functional interconnection of the components in living cells. Altogether, our findings will reveal the mechanism by which TGN sorts and targets proteins to their respective destination, a process that is essential for cell compartmentation, tissue organization, and function.

项目摘要 对可溶性蛋白质进行分选以运输到细胞内的隔室和从细胞中分泌是必不可少的 维持细胞和组织的动态平衡。细胞内蛋白质分布的一个主要调节因素是反式高尔基体 将分泌蛋白质分选到特定载体以将其运送到最终目的地的网络(TGN)。 通过甘露糖-6-磷酸受体对TGN上的溶酶体水解酶进行分选已得到很好的证实。 然而，对分泌蛋白的分选机制仍然知之甚少。最近发现的 在TGN对分泌货物的基于钙的分类开始揭示细胞 分类来自高尔基居民的秘密货物和运往其他蜂窝隔间的货物。这 基于钙离子的分选涉及细胞质肌动蛋白细胞骨架，它通过膜锚定的钙离子 ATPase SPCA1和腔钙结合蛋白Cab45是位于 TGN.然而，规范这些组件之间相互作用的基本机制并不是 安全。我们采取跨学科和高度创新的方法来解决这一根本问题 重要性。我们的目标包括在模型膜中对这些成分进行体外重组，以确定 对秘密货物进行分类所需的最小机械。现代基因组编辑技术在中国 与荧光和超分辨率显微镜相结合的人类细胞将提供对 活细胞中组件的功能互连。总之，我们的发现将揭示 TGN将蛋白质分类并定位于其各自目的地的机制，这一过程是 对细胞分隔、组织组织和功能是必不可少的。