CAREER: Towards Synthetic Cytoplasm: Volume Exclusion and Aqueous Phase Separation in Giant Vesicles

这项授予宾夕法尼亚州立大学克里斯汀·基廷教授的职业奖项得到了化学部的分析和表面化学项目以及分子和细胞生物科学部的细胞系统的支持。在这个项目中，将研究包含在巨大的单层囊泡中的双水相系统的性质。要解决的问题包括蛋白质和其他大分子在两个相之间的分配，以及这种分配如何受到囊泡内胶囊的影响。分配可以遵循Michaelis-Menton，或者可以显示S型，即“开关状”响应曲线，后者提供独特的囊泡特性。需要研究的一种性质是囊泡运动，这种运动基于细胞骨架蛋白在这些“合成细胞”内的可逆聚合，驱动力由温度和/或化学梯度提供。该项目的总体目标是阐明细胞所使用的功能结构，这一知识随后可以应用于非生物纳米结构。该项目描述了一个全面的教育和推广计划，包括通过高中生物教师向高中生推广。高中教师已经在暑假期间在PI的实验室工作了几个月，一名新教师将用这个奖项支持一个夏天。这些教师使用最先进的仪器，并在开发适用于高中实验室的生物传感器和细胞结构的新实验方面发挥了关键作用。此外，国际生物研究所将开发一门新的研究生水平的生物识别课程。

This CAREER award to Professor Christine Keating of Pennsylvania State University is supported by the Analytical and Surface Chemistry Program in the Chemistry Division and by Cellular Systems in the Division of Molecular and Cellular Biosciences. In this project, the properties of aqueous two-phase systems will be studies as contained within giant unilamellar vesicles. Questions to be addressed include the partitioning of proteins and other macromolecules between the two phases, and how this partitioning is affected by encapsulation within the vesicle. The partitioning could either follow Michaelis-Menton or may show a sigmoidal, i.e., "switchlike" response curve, with the latter giving access to unique vesicle properties. One property to be examined is vesicle locomotion based on the reversible polymerization of cytoskeletal proteins within these "synthetic cells" with the driving force provided by a temperature and/or chemical gradient. The overall goal of the project is elucidation of the functional architectures used by cells, a knowledge which can then be applied to abiotic nanostructures.A comprehensive education and outreach plan is described, including outreach to high school students through a high school biology teacher. High school teachers have already worked in the PI's lab during summer months and a new teacher is to be supported for one summer with this award. These teachers have worked with state-of-the-art instrumentation and have been key in development of new experiments with biosensors and cell structure appropriate for the high school laboratory. In addition, the PI will develop a new graduate-level course on biorecognition.