STRUCTURAL MECHANISMS OF GTPASE AND PHOPHOINOSTIDE REGULATED TRAFFICKING

GTPase 和磷酸肌苷调控贩运的结构机制

• 批准号: 7299617
• 负责人: David G Lambright
• 金额: $ 57.87万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-10 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7299617
• 关键词: 1-Phosphatidylinositol 3-Kinase; Adipose tissue; Affinity; Amino Acid Substitution; Arts; Binding; Binding Sites; Biochemical; C-terminal; Cell Surface Receptors; Cell membrane; Cells; Collaborations; Complex; Data; Defect; Disease; Endocytosis; Epidermal Growth Factor; Event; Family; Funding; GTPase-Activating Proteins; Glucose Transporter; Growth; Growth Factor; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate Phosphohydrolases; Homeostasis; Image; Insulin; Insulin Receptor; Length; Lipids; Malignant Neoplasms; Maps; Mediating; Membrane; Membrane Proteins; Metabolic; Metabolism; Molecular; Muscle Cells; Mutation; N-terminal; Non-Insulin-Dependent Diabetes Mellitus; Nucleotides; PH Domain; Phosphatidylinositols; Play; Principal Investigator; Protein Family; Protein Isoforms; Proteins; Public Health; Reagent; Receptor Signaling; Recycling; Regulation; Role; Signal Pathway; Signal Transduction; Specificity; Structure; System; Testing; Therapeutic Intervention; Transferrin Receptor; Ubiquitin; base; cancer type; cytohesin-1; molecular assembly/self assembly; novel; programs; rab GTP-Binding Proteins; ras Proteins; receptor; response; spatiotemporal; src Homology Region 2 Domain; stoichiometry; trafficking

Objective and Aims for Project 3 The internalization and recycling of cell surface receptors, glucose transporters, and other plasma membrane proteins is regulated by phosphoinositide metabolism as well as GTPases of the Rab and Arf families. Defects in these regulatory mechanisms have been implicated in a variety of disease states including cancer and type II diabetes. The long term objective of Project 3 is to characterize poorly understood mechanisms that integrate phosphoinositide signaling and Rab/Arf GTPase activation with the dynamic molecular assemblies that mediate/regulate trafficking events at the plasma membrane and within the endosomal system. To achieve this objective, we will combine crystallographic, biochemical, biophysical, and mutational studies on purified complexes with complementary structure-function analyses in collaboration with Projects 1, 2, 4, and the imaging core. Building on the observations of the previous funding period and extensive new preliminary data, we will: (Aim 1) examine the structural determinants of autoregulation and phosphoinositide dependent membrane targeting in the Grp1 family of Arf GTPase exchange factors; (Aim 2) investigate the mechanism of autoregulation, membrane targeting, and Rab activation by the Rabex-5/Rabaptin-5 complex and Rin1;and (Aim 3) explore the structural bases for regulation of endocytic recycling by phosphoinositide dependent and independent complexes of EHD proteins with Rabenosyn-5, EHBP1, and FIP2. Relevance to Public Health Both type II diabetes and cancer involve defects in the mechanisms by which cell surface receptors for insulin and growth factors communicate with the molecular machinery that controls cell metabolism and growth. By studying these mechanisms comprehensively at the structural, molecular, and cellular levels through collaborative projects the combine the expertise of structural, molecular, and cell biologists we hope to identify novel mechanism-based strategies that can be exploited for therapeutic intervention.

项目3的目标和目标 细胞表面受体、葡萄糖转运体和其他血浆的内化和再循环 膜蛋白受肌醇磷脂代谢以及Rab和Arf的GTP酶的调节 家人。这些调节机制的缺陷被认为与多种疾病状态有关 包括癌症和II型糖尿病。项目3的长期目标是描述糟糕的特征 了解肌醇磷脂信号和Rab/Arf GTP酶激活与 在质膜和膜内调节转运事件的动态分子集合体 内吞体系统。为了实现这一目标，我们将结合结晶学、生化、 用互补结构-功能分析对纯化的配合物进行生物物理和突变研究 与项目1、2、4和成像核心进行协作。建立在以前观察到的基础上 资金期限和广泛的新的初步数据，我们将：(目标1)审查结构性决定因素 Arf GTP酶Grp1家族的自身调节和肌醇磷脂依赖的膜靶向 交换因子；(目的2)研究自动调节、膜靶向和RAB的机制 由Rabex-5/Rabaptin-5复合体和Rin1激活；以及(目标3)探索Rabex-5/Rabaptin-5复合体和Rin1的结构基础 EHD磷脂酰肌醇依赖和非依赖复合体对细胞内循环的调节 含有Rabenosyn-5、EHBP1和FIP2的蛋白质。 与公共卫生的相关性 II型糖尿病和癌症都涉及细胞表面受体 胰岛素和生长因子与控制细胞新陈代谢和 成长。通过在结构、分子和细胞水平上全面研究这些机制 通过合作项目，我们希望将结构、分子和细胞生物学家的专业知识结合在一起 确定可用于治疗干预的新的基于机制的策略。