Intracellular Receptor Trafficking

细胞内受体贩运

• 批准号: 7560913
• 负责人: MANUEL A BARBIERI
• 金额: $ 26.51万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-09 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7560913
• 关键词: Address; Affect; American; Area; Binding; Biochemical; Biological Assay; Cell Surface Receptors; Cell membrane; Cells; Clathrin; Cognitive; Collaborations; Coupled; Data; Development; Diabetes Mellitus; Disease; Early Endosome; Educational workshop; Endocytosis; Endosomes; Epidermal Growth Factor; Event; Funding; Goals; Grant; Growth Factor; Guanosine Triphosphate Phosphohydrolases; In Vitro; Insulin Receptor; Insulin Signaling Pathway; Laboratories; Lead; Link; Malignant Neoplasms; Manuscripts; Mediating; Membrane Protein Traffic; Mentors; Metabolic; Methodology; Minority; Modeling; Modification; Molecular; Molecular and Cellular Biology; Mutagenesis; Paper; Pathway interactions; Peer Review; Physiological; Physiological Processes; Physiology; Play; Polishes; Postdoctoral Fellow; Principal Investigator; Proteins; Proteomics; Publishing; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Regulation; Research; Resources; Risk; Risk Factors; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Specificity; Students; Time; Tissues; Training; Travel; United States National Institutes of Health; Vesicle; Work; Writing; attenuation; cell growth; glucose metabolism; human disease; hypertensive heart disease; insulin signaling; interdisciplinary approach; interest; macromolecule; mortality; novel; protein function; receptor; receptor internalization; receptor mediated endocytosis; reconstitution; therapeutic target; trafficking

DESCRIPTION (provided by applicant): The long-term goal is to employ a multidisciplinary approach to gain a thorough understanding of the molecular mechanisms by which Rin1 regulates both insulin receptor (IR)-mediated trafficking and signaling. Elucidation of the mechanisms by which Rin1 regulates IR signaling and trafficking may serve as a model for tyrosine receptor kinases and other receptor signal transducing proteins. Each of these aims is significant because the mechanistic and physiologic functions of the Rin1 molecules are unknown, as are the mechanisms used to regulate Rin1 protein function. Thus, we have a unique opportunity to determine whether such effects are sufficient to alter the downstream effect of the IR signaling pathways, a question of considerable interest to the physiology of the insulin-signaling field. Therefore, the strength of this project is its ability to reveal new signaling regulatory mechanisms that are directly relevant to an important physiologic process and which may be defective in some cases of human disease. Accordingly, our objectives are to: 1) Determine how Rin1 is coupled to the activated insulin receptor. We will elucidate the specificity of this Rin1-insulin receptor interaction via mutagenesis and biochemical binding assays. 2) Determine how Rin1 is linked to the endocytic trafficking of the activated insulin receptor. We will address the mechanism of Rin1 function during internalization of insulin receptor through the activation of Rab5 GTPase. 3) Determine how Rin1 is integrated to the insulin receptor signaling transduction pathways. We postulate that Rin1 is required for insulin receptor signaling, which may favor one or more signal transduction pathways. 4) Determine the role of Rin1 in early endosome fusion. We will reconstitute and characterize Rin1-dependent endosome fusion by using a novel in vitro endosome fusion assay. Delineating the cellular and molecular biology of Rin1 will expand our understanding of the role of receptor internalization and membrane trafficking in signal transduction and will also help to define therapeutic targets for selectively modulating signal transduction via tyrosine kinase receptors. Diabetes affects an estimated 20 million Americans, making this disease one of the leading mortality risk factors due its association with greatly increased risk of heart disease, hypertension and cancer. This proposal is directly relevant to the goal of establishing molecular factors that lead to affecting insulin receptor signaling and action. It addresses whether functional modification of insulin signaling pathways, one that is predominant in metabolic tissues --impaired glucose metabolism-- and another being a growth factor-like pathway --cell growth ad proliferation-- lead to diabetes by impaired insulin receptor trafficking and signaling, a key question in the field. In this project, we have a unique opportunity to address this question by elucidating the molecular mechanism by which Ras interference 1 regulates insulin receptor-signaling and intracellular trafficking and may also serve as a model for tyrosine receptor kinases. The key feature of this project is its ability to reveal new signaling regulatory mechanisms that are relevant to an important physiologic process, such as diabetes.

描述（由申请人提供）：长期目标是采用多学科方法，全面了解Rin 1调节胰岛素受体（IR）介导的运输和信号传导的分子机制。阐明Rin 1调节IR信号传导和运输的机制可以作为酪氨酸受体激酶和其他受体信号转导蛋白的模型。这些目标中的每一个都是重要的，因为Rin 1分子的机制和生理功能是未知的，用于调节Rin 1蛋白功能的机制也是未知的。因此，我们有一个独特的机会，以确定这种影响是否足以改变IR信号通路的下游效应，一个相当感兴趣的问题，胰岛素信号领域的生理学。因此，该项目的优势在于它能够揭示与重要生理过程直接相关的新的信号调节机制，这些机制在某些人类疾病中可能存在缺陷。因此，我们的目标是：1）确定Rin 1是如何耦合到激活的胰岛素受体。我们将通过诱变和生化结合试验阐明Rin 1-胰岛素受体相互作用的特异性。2)确定Rin 1是如何与激活的胰岛素受体的内吞运输相联系的。我们将通过Rab 5 GT3的激活来阐明Rin 1在胰岛素受体内化过程中的作用机制。3)确定Rin 1如何整合到胰岛素受体信号转导通路中。我们推测Rin 1是胰岛素受体信号传导所必需的，这可能有利于一个或多个信号转导途径。4)确定Rin 1在早期内体融合中的作用。我们将使用一种新的体外核内体融合试验重建和表征Rin 1依赖性核内体融合。描绘Rin 1的细胞和分子生物学将扩大我们对受体内化和膜运输在信号转导中的作用的理解，也将有助于确定通过酪氨酸激酶受体选择性调节信号转导的治疗靶点。据估计，糖尿病影响着2000万美国人，使这种疾病成为主要的死亡风险因素之一，因为它与心脏病、高血压和癌症的风险大大增加有关。这一提议与建立导致影响胰岛素受体信号传导和作用的分子因子的目标直接相关。它解决了胰岛素信号传导途径的功能修饰，一个是主要的代谢组织-受损的葡萄糖代谢-另一个是生长因子样途径-细胞生长和增殖-通过受损的胰岛素受体运输和信号传导导致糖尿病，这是该领域的一个关键问题。 在这个项目中，我们有一个独特的机会来解决这个问题，阐明Ras干扰1调节胰岛素受体信号和细胞内运输的分子机制，也可以作为酪氨酸受体激酶的模型。该项目的主要特点是能够揭示与重要生理过程（如糖尿病）相关的新信号调节机制。