CAREER: Optically Controlled Protein Proximity Labelling

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With support from the Chemistry of Life Processes Program in the Division of Chemistry, Dr. Michael Taylor at the University of Wyoming will develop new, light-promoted chemical reactions that enable study of the biological role of the mitochondrial translocator protein. Proteins found within a cell form complex networks with neighboring proteins to help control critical cellular processes. In these networks, proteins interact and communicate with each other by packing very closely together in specific orientations. These networks often change rapidly in response to external stimuli or because of cellular dysfunction, making them difficult to study in their natural environment. In this project, Dr. Taylor will study protein networks by developing a new chemical reaction that labels a protein of interest with an unnatural covalent bond to a chemical probe. The labeling chemistry that is developed in this project is controlled by application of a specific color of light. The project exploits this requirement through precision molecular design of the probes, which is expected to enable iterative protein labeling sequences that are achieved by simply altering the color of light used to activate the reaction. This would allow for the study of not only a specific protein itself, but also how the target protein networks with neighboring proteins, and how these networks change in real time. Integrated educational objectives focus on increasing awareness of and enthusiasm for science-technology-engineering and mathematics (STEM) in the state of Wyoming through two key activities: (1) piloting on-line, interactive curricula at the middle school level, which emphasize how biology and chemistry are inherently linked; and (2) using the chemical reactions that are developed in this project, that employ aesthetically pleasing fluorescent molecules, as a basis for interactive demonstrations for local Girl Scout troops. With this CAREER Award, the Chemistry of Life Processes Program is funding Dr. Michael Taylor at the University of Wyoming to develop new, light-promoted chemical reactions that are applied to the study of the biological role of the mitochondrial translocator protein, tryptophan-rich sensory protein (TSPO). TSPO has proven to be a challenging protein to study with canonical experimental workflows, and thus, its proposed roles in cellular function remain controversial. Dr. Taylor seeks to address this challenge by developing a bioconjugation reaction that combines multiple selectivity factors, including light activation, to chemically label TSPO with high precision. Dr. Taylor aims to exploit the modular tunability of the chemical systems employed herein to develop a suite of reagents that are activated over a wide array of wavelengths and can achieve protein labeling through different mechanisms. This new approach opens up the possibility of using optically orchestrated, iterative protein labeling sequences to label both TSPO itself, as well as neighboring proteins. This, in turn, could enable the construction of a three-dimensional rendering of the biological environment surrounding TSPO, which would lend unique insight into the biological role of TSPO. This CAREER Award provides the support to develop, understand, and apply a suite of chemical reagents and reactions that allow for the study of TSPO in a cellular environment. The integration of education and research is achieved through two specific educational activities designed to enhance enthusiasm and awareness of STEM by developing interactive STEM curricula for middle school students and implementing interactive demonstrations for local Girl Scout troops.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.