Rab11-FIPs interact to define distinct functional nodes in the recycling system

Rab11-FIP 相互作用以定义回收系统中不同的功能节点

• 批准号: 7481131
• 负责人: Jenny Schafer
• 金额: $ 4.96万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7481131
• 关键词: Alzheimer' s Disease; Binding; Cell Surface Proteins; Cell membrane; Cell physiology; Cell surface; Cells; Class; Complex; Cystic Fibrosis; Data; Diabetes Insipidus; Disease; Docking; Ensure; Environment; Family; Fluorescence Resonance Energy Transfer; Heterodimerization; Homodimerization; In Situ; Lead; Location; Maintenance; Malignant Neoplasms; Mammalian Cell; Microscopy; Molecular Conformation; Monomeric GTP-Binding Proteins; Myosin ATPase; Pattern; Process; Proteins; Recycling; Regulation; Role; Specificity; System; Testing; Time; Transport Vesicles; Yeasts; cell type; intermolecular interaction; intracellular protein transport; protein localization location; protein transport; rab GTP-Binding Proteins; rab11 protein; receptor; trafficking; yeast two hybrid system

DESCRIPTION (provided by applicant): Protein trafficking and transport is a crucial aspect of cellular function because cells interact with the environment by internalization and release of proteins from the cell surface. Aberrant processing or transport of cell surface proteins contributes to diseases such as diabetes insipidus, cystic fibrosis, and Alzheimer's disease. While the particular cargo such as channels and receptors is unique to different cell types, the process of plasma membrane recycling is highly conserved among mammalian cells and regulated by a common set of trafficking proteins including the Rab small GTPases. Rab11 is a ubiquitously expressed small GTPase involved in recycling that interacts with the Rab11-Family Interacting Proteins (FIPs). Although the functions of FIPs are not fully understood, they may in part define distinct subdomains within the recycling system. Furthermore, preliminary data suggest that FIP2 exists in different conformations in different subcellular localizations. These data lead to the hypothesis that discrete oligomers of FIPs interacting with Rab11 allow for temporal and spatial specificity of Rab11 and FIP function within the recycling system. Two aims are proposed to test this idea. First, homodimerization and heterodimerization capabilities of FIPs will be assessed by yeast two hybrid, FRET microscopy, and protein colocalization as a way to understand FIP function within the recycling system. Second, FIP2 conformation will be studied with regards to intramolecular and intermolecular interactions within the recycling system. By elucidating the composition of FIP complexes and their subcellular localizations, the importance of Rab11 and FIPs as general regulators of plasma membrane recycling may be uncovered. SIGNIFICANCE: Proper protein transport and localization are crucial to maintenance of normal cellular function. Diseases such as cystic fibrosis, diabetes insipidus, and Alzheimer's disease result from mislocalization or aberrant trafficking of vital proteins. This proposal aims to increase the understanding of the basic mechanism of protein trafficking that is common to all cell types by studying an important trafficking protein, Rab11, and its interacting proteins.

描述（由申请人提供）：蛋白质运输和运输是细胞功能的关键方面，因为细胞通过内在化和从细胞表面释放蛋白质与环境相互作用。细胞表面蛋白的异常加工或运输会导致糖尿病，囊性纤维化和阿尔茨海默氏病等疾病。尽管特定的货物（例如通道和受体）是不同细胞类型的独有的，但质膜回收的过程在哺乳动物细胞中高度保守，并由一组常见的运输蛋白（包括Rab Small GTPase）调节。 Rab11是一种与Rab11家族相互作用蛋白相互作用（FIPS）相互作用的普遍表达的小GTPase。尽管FIP的功能尚未完全理解，但它们可能部分地定义了回收系统中不同的子域。此外，初步数据表明，在不同的亚细胞定位中，FIP2存在不同的构象。这些数据导致了以下假设：FIPS与RAB11相互作用的离散低聚物允许在回收系统中具有RAB11和FIP功能的时间和空间特异性。提出了两个目标来测试这一想法。首先，FIP的同构二聚化和异二聚化能力将通过酵母两杂交，FRET显微镜和蛋白质共定位，作为了解回收系统中FIP功能的一种方式。其次，将对回收系统内分子内和分子间相互作用进行研究。通过阐明FIP复合物的组成及其亚细胞局部化，可以发现Rab11和FIP作为质膜回收的一般调节剂的重要性。 意义：正确的蛋白质转运和定位对于维持正常细胞功能至关重要。囊性纤维化，糖尿病和阿尔茨海默氏病等疾病是由于非定位或对重要蛋白质的异常贩运而引起的。该提案旨在通过研究重要的运输蛋白Rab11及其相互作用蛋白质来增加对蛋白质运输的基本机制的理解。