Imaging the early events in membrane receptor signaling

项目摘要 了解形成有效细胞反应的分子机制是一个 生物学的基本问题。虽然我们已经对这一系列事件有了很多了解， 膜受体-配体结合，在我们的理解如何有一个根本的差距 蛋白质动力学促进信号传导。我的研究计划的长期目标是 了解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。这项提案的目的是量化蛋白质 在膜相关信号传导的早期事件期间的动态。我们的核心假设是 蛋白质相互作用的持续时间调节信号的结果。的理由 为了理解信号转导， 确定启动的生化事件的序列、寿命和亚细胞定位， 发信号。我们使用独特和创新的成像技术提供定量信息 早期信号事件的动力学，不能使用传统的生化 （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，在一个综合的方法，以提供 蛋白质-蛋白质动力学如何调节酪氨酸激酶信号传导的全面图片 生长因子受体和免疫受体的下游。拟议的研究是 重要的是，我们将获得的定量信息还没有直接 测量之前，并将带来新的观点，细胞生物学过程，无论是在正常 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，信号伙伴相互作用和下游信号事件，我们将填补 我们细胞信号模型中的关键时空缺口。

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.