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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 G蛋白、它们的偶联受体和潜在的通路是药物开发的目标，目前所有批准的治疗药物对这些系统的影响程度都超过50%。利用荧光技术更好地了解G蛋白如何响应细胞外信号，安装响应，以及这种高度保守的系统的潜在调节，有助于传感器的健壮性和动态范围。双光子显微镜将被用来定量荧光标记蛋白的位置和动力学，如受体，G蛋白和下游效应器。FRET和FCS/ICS将被用来了解系统的分子动力学、蛋白质数量、扩散速率和扩散向量，并建立可用于建立计算模型的数据体。交配过程的其他方面也将被观察到，例如膜的流动性和组成及其随时间的变化，蛋白质招募和内化的定位，以及蛋白质与这些现象的共存。

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.