Spectroscopic Studies of EGF-Receptor Interactions

EGF-受体相互作用的光谱研究

• 批准号: 6905530
• 负责人: ALBERT H BETH
• 金额: $ 29.56万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6905530
• 关键词: SDS polyacrylamide gel electrophoresis; binding sites; biological signal transduction; cell line; epidermal growth factor; flow cytometry; fluorescence spectrometry; glycosylation; growth factor receptors; high performance liquid chromatography; intermolecular interaction; ligands; matrix assisted laser desorption ionization; membrane proteins; mitogen activated protein kinase; peptide hormone; polymerase chain reaction; protein tyrosine kinase; receptor binding; stop flow technique; surface plasmon resonance

DESCRIPTION (provided by applicant): This project combines resources of two established laboratories in membrane protein biophysics and biochemistry to study details of the intermolecular interactions of the epidermal growth factor (EGF) receptor with its ligands and with other receptors of the ErbB family of receptor kinases. The ErbB family of receptors, and, in particular, the EGF receptor, are among the most widely studied polypeptide hormone receptors because of their importance in normal development, wound healing, and, when deregulated by mutation or over-expression, neoplasia. Nonetheless, fundamental questions concerning the mechanisms of signal transduction triggered by the binding of the EGF family of ligands to the ErbB family of receptors remain unresolved. The long term goal of this project is to develop an in-depth understanding of the earliest events in the signal transduction pathway that occur when EGF family ligands bind to their respective receptors. The specific aims for this budget period will exploit well characterized fluorescent ligands, new cell lines that have been shown to withstand turbulent stoppedflow rapid mixing and that express selected ErbB family receptors in the absence of endogenous ErbB receptors, as well as a custom stopped-flow fluorometer specifically designed and optimized for these studies. Studies will address hypotheses a) that expression of other ErbB family members affects the proportion of EGF receptors in high and low affinity states, b) that ligands recognized by ErbB3 or ErbB4 can modulate the binding of EGF to its receptor when the receptors are present in the same cell, c) that the dynamic on and off rates for receptor binding are important parameters for determining the mitogenic activity ofpeptide hormones, and d) that heterogeniety in glycosylation can affect receptor affinity for EGF. Studies will also test competing hypotheses concerning the interactions between the EGF receptor and ErbB2: that EGF receptor-ErbB2 dimers form directly when EGF binds to the receptor, or that EGF receptors dimerize on binding EGF and subsequently recruit ErbB2.

描述（由申请人提供）：该项目结合了膜蛋白生物物理学和生物化学领域两个已建立的实验室的资源，研究表皮生长因子（EGF）受体与其配体以及与受体激酶 ErbB 家族的其他受体的分子间相互作用的细节。 ErbB 受体家族，特别是 EGF 受体，是研究最广泛的多肽激素受体之一，因为它们在正常发育、伤口愈合以及因突变或过度表达而失调时的肿瘤形成中发挥着重要作用。尽管如此，关于 EGF 配体家族与 ErbB 受体家族结合引发的信号转导机制的基本问题仍未解决。该项目的长期目标是深入了解 EGF 家族配体与其各自受体结合时发生的信号转导途径中的最早事件。本预算期的具体目标将利用特征明确的荧光配体、已证明能够承受湍流停流快速混合并在缺乏内源性 ErbB 受体的情况下表达选定的 ErbB 家族受体的新细胞系，以及专门为这些研究设计和优化的定制停流荧光计。研究将提出以下假设：a) 其他 ErbB 家族成员的表达影响高亲和力状态和低亲和力状态下 EGF 受体的比例，b) 当受体存在于同一细胞中时，ErbB3 或 ErbB4 识别的配体可以调节 EGF 与其受体的结合，c) 受体结合的动态开启和关闭速率是确定肽激素促有丝分裂活性的重要参数， d) 糖基化的异质性会影响受体对 EGF 的亲和力。研究还将测试有关 EGF 受体和 ErbB2 之间相互作用的竞争假设：当 EGF 与受体结合时，EGF 受体-ErbB2 二聚体直接形成，或者 EGF 受体在结合 EGF 时形成二聚体，随后招募 ErbB2。