PROTEIN NETWORKS AND FORCE GENERATION IN YEAST ENDOCYTOSIS

酵母内吞作用中的蛋白质网络和力产生

基本信息

批准号：
8848850
负责人：
ANDERS E CARLSSON
金额：
$ 28.98万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-15 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8848850
关键词：
Actins Address Alzheimer&apos s Disease Area Automobile Driving Binding Biological Assay Biological Models Biomedical Engineering Biophysical Process Capsid Proteins Cell Surface Receptors Cell membrane Cell physiology Cells Cellular biology Chemistry Clathrin Collaborations Complex Computer Simulation Coupled Disease Elements Endocytic Vesicle Endocytosis Equation F-Actin Feedback Filopodia Gene Proteins Generations Geometry Growth Health Human Image In Vitro Lateral Malignant Neoplasms Mechanics Mediating Membrane Methods Microfilaments Modeling Molecular Molecular Genetics Motor Activity Mutation Myosin ATPase Output Peptides Phase Physics Play Process Production Protein Dynamics Proteins Public Health Research Project Grants Resolution Role Side Spatial Distribution Staging Tertiary Protein Structure Testing Time Toxin Universities Washington Work Yeasts base cell motility cell type driving force extracellular in vivo light microscopy mathematical methods mathematical model medical schools polymerization pressure professor receptor research study segregation simulation theories tool

项目摘要

DESCRIPTION: The project aims to elucidate key biophysical mechanisms underlying clathrin-mediated endocytosis in yeast. Endocytosis involves a network of over sixty proteins, which exert forces that deform the cell membrane. The protein actin, whose polymerization is controlled by upstream proteins called nucleation-promoting-factors, plays a major role in generating force and controlling protein assembly. We will explore the functionality of the endocytic protein network, and the mechanisms by which force is generated, through mathematical modeling, simulation and quantitative experimental studies. The mathematical methods include stochastic simulation of actin polymerization, and a finite- element approach to membrane deformation. The experimental methods include actin polymerization assays and high-resolution imaging of key endocytic proteins. The experimental outputs will be compared directly to the theoretical predictions. The combination of theory and experiment will explain how multiple nucleation promoting factors regulate actin polymerization; how different nucleation-promoting factors become segregated to different parts of the endocytic tubule; and how the spatial distribution of forces leading to endocytosis is regulated by actin and curvature-generating proteins. Because of its small spatial scale, and the limited number of proteins involved, clathrin- mediated endocytosis is a model process for understanding the function of intracellular mechanochemical networks - networks where chemistry is coupled bidirectionally to force generation and membrane deformation. Therefore our understanding of endocytosis will help us understand the more complex mechanochemical networks in cells, such as those driving ruffling, lamellipodium or filopodium formation, and cell migration.

描述：该项目旨在阐明酵母中网状蛋白介导的内吞作用的关键生物物理机制。内吞作用涉及一个超过60种蛋白质的网络，该网络施加了使细胞膜变形的力。蛋白质肌动蛋白的聚合由称为成核促进因子的上游蛋白控制，在产生力和控制蛋白质组装方面起着主要作用。我们将探索内吞蛋白网络的功能，以及通过数学建模，仿真和定量实验研究生成力的机制。数学方法包括肌动蛋白聚合的随机模拟，以及膜变形的有限元方法。实验方法包括肌动蛋白聚合测定和关键内吞蛋白的高分辨率成像。实验输出将直接与理论预测进行比较。理论和实验的结合将解释多种成核如何促进肌动蛋白聚合。如何将不同的启动因子隔离到内吞小管的不同部位；以及如何通过肌动蛋白和曲率生成的蛋白来调节导致内吞作用的力的空间分布。由于其较小的空间尺度和涉及的蛋白质数量有限，网格蛋白介导的内吞作用是理解细胞内机械化学网络的功能的模型过程 - 在该网络上，化学上的化学性质是双向耦合以产生力和膜变形的。因此，我们对内吞作用的理解将有助于我们了解细胞中更复杂的机械化学网络，例如那些驾驶荷叶边，层状脂蛋白或丝状地位形成以及细胞迁移。