QUANTITATIVE ANALYSIS OF G-PROTEINS & COUPLED RECEPTORS IN YEAST MATING

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 G蛋白、其偶联受体和潜在途径作为药物开发的靶点引起了极大的兴趣，目前所有批准的治疗剂对这些系统的影响水平超过50%。利用荧光技术更好地了解G蛋白如何响应细胞外信号，建立响应，以及这些系统的潜在调控，这是高度保守的，有助于传感器的鲁棒性和动态范围。双光子显微镜将用于定量荧光标记蛋白质的位置和动态，如受体，G蛋白和下游效应物。FRET和FCS/ICS将用于了解系统的分子动力学，蛋白质数量，扩散速率和扩散矢量，并建立一个数据库，然后可以用于制作计算模型。还将观察交配过程的其他方面，例如膜流动性和组成及其随时间的变化，蛋白质募集和内化的定位，以及蛋白质与这些现象的共定位。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. G-proteins, their coupled receptors, and underlying pathways are of great interest as targets for pharmaceutical development, with current levels of all approved therapeutic agents affecting these systems at greater than 50%. Using florescence techniques a greater understanding how G-proteins respond to extra cellular signals, mount a response, and the underlying regulation of such systems, which are highly conserved, contributes to the robustness and dynamic range of the sensors.Two-photon microscopy will be utilized to quantitate the location and dynamics of fluorescently labeled proteins, such as the receptor, g-proteins and downstream effectors. FRET, and FCS/ICS will be used to understand the molecular dynamics of the system, protein number, diffusion rate, and diffusion vector, and build a body of data that can then be used to make computational models. Other aspects of the mating process will also be observed, such as membrane fluidity and composition and it changes over time, localization of protein recruitment and internalization, and co-localization of proteins with these phenomena.