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

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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.