Imaging the early events in membrane receptor signaling

项目摘要 理解形成有效细胞反应的分子机制是一个 生物学中的基本问题。虽然人们对由 膜受体与配体的结合，在我们对如何结合的理解上有一个根本性的差距 蛋白质动力学有助于信号传递。我的研究计划的长期目标是 了解蛋白质-蛋白质相互作用的动态和随机行为是如何整合的 以产生有效的信号响应。这项提议的目标是量化蛋白质 膜相关信号早期事件中的动力学。我们的中心假设是 蛋白质-蛋白质相互作用的持续时间调节信号结果。其基本原理是 对于拟议的研究来说，为了理解信号转导，关键是 确定启动生化事件的序列、生命周期和亚细胞定位 发信号。我们使用独特和创新的成像技术来提供定量信息 关于用传统生化方法无法获得的早期信号事件的动力学 (基于人口的)技术。我们将应用我们独特的工具箱，包括最先进的 显微镜方法、生物物理和功能读出，在综合方法中提供 蛋白质-蛋白质动力学如何调节酪氨酸激酶信号的综合图景 生长因子受体和免疫受体下游。拟议的研究是 意义重大，因为我们将获得的定量信息并没有直接 以前测量过，并将为细胞生物学过程带来新的视角，无论是在正常情况下 和疾病状态。通过将细胞表面受体的激活状态与它们的 动态、信令合作伙伴相互作用和下行信令事件，我们将填写 我们的细胞信号模型中的关键时空间隙。

Project Summary Understanding the molecular mechanisms that shape an effective cellular response is a fundamental question in biology. While much is known about the chain of events initiated by membrane receptor-ligand binding, there is a fundamental gap in our understanding of how protein dynamics facilitate signaling. The long-term goal of my research program is to understand how the dynamic and stochastic behavior of protein-protein interactions is integrated to produce an efficient signaling response. The objective of this proposal is to quantify protein dynamics during the early events of membrane-associated signaling. Our central hypothesis is that the duration of protein-protein interactions modulates the signaling outcome. The rationale for the proposed research is that in order to understand signal transduction, it is critical to determine the sequence, lifetime and sub-cellular localization of biochemical events that initiate signaling. We use unique and innovative imaging techniques to provide quantitative information on the dynamics of early signaling events that cannot be obtained using traditional biochemical (population-based) techniques. We will apply our unique tool box, including state-of-the-art microscopy methods, biophysical and functional read-outs, in an integrated approach to provide a comprehensive picture of how protein-protein dynamics regulate tyrosine kinase signaling downstream of growth factor receptors and immunoreceptors. The proposed research is significant because the quantitative information that we will obtain has not been directly measured before and will bring new perspectives to cell biological processes, both in normal and disease states. By connecting the activation state of cell surface receptors with their dynamics, signaling partner interplay and downstream signaling events, we will are filling in critical spatiotemporal gaps in our models of cell signaling.