CAREER: Optically Controlled Protein Proximity Labelling

在化学系生命过程化学项目的支持下，怀俄明大学的迈克尔·泰勒博士将开发新的光促进化学反应，使研究线粒体转位蛋白的生物学作用成为可能。细胞内发现的蛋白质与邻近的蛋白质形成复杂的网络，帮助控制关键的细胞过程。在这些网络中，蛋白质通过在特定方向上非常紧密地堆积在一起来相互作用和交流。这些网络经常因外界刺激或细胞功能障碍而迅速变化，使它们很难在自然环境中进行研究。在这个项目中，泰勒博士将通过开发一种新的化学反应来研究蛋白质网络，这种化学反应将目标蛋白质标记为与化学探针不自然的共价键。在这个项目中开发的标记化学是通过应用特定颜色的光来控制的。该项目通过对探针进行精确的分子设计来利用这一要求，预计这将使重复的蛋白质标记序列能够通过简单地改变用于激活反应的光的颜色来实现。这不仅可以研究特定蛋白质本身，还可以研究目标蛋白质如何与邻近蛋白质网络，以及这些网络是如何实时变化的。综合教育目标侧重于通过两项关键活动提高怀俄明州对科学-技术-工程与数学(STEM)的认识和热情：(1)在中学层面试行在线互动课程，强调生物和化学是如何内在联系的；(2)利用该项目开发的化学反应，使用美观的荧光分子，作为当地女童子军互动演示的基础。凭借这一职业奖，生命过程化学计划资助怀俄明大学的Michael Taylor博士开发新的光促进化学反应，用于研究线粒体转位蛋白-富含色氨酸的感觉蛋白(TSPO)的生物学作用。TSPO已被证明是一个具有挑战性的蛋白质，通过规范的实验工作流程进行研究，因此，它在细胞功能中的拟议作用仍然存在争议。泰勒博士试图通过开发一种生物偶联反应来应对这一挑战，该反应结合了包括光激活在内的多种选择性因素，以高精度的化学标记TSPO。泰勒博士的目标是利用本文使用的化学体系的模块化可调控性来开发一套试剂，这些试剂可以在广泛的波长范围内激活，并可以通过不同的机制实现蛋白质标记。这种新的方法打开了使用光学编排的、迭代的蛋白质标记序列来标记TSPO本身以及邻近蛋白质的可能性。这反过来又可以构建TSPO周围生物环境的三维渲染，这将为TSPO的生物学作用提供独特的见解。该职业奖为开发、理解和应用一套化学试剂和反应提供支持，这些试剂和反应允许在细胞环境中研究TSPO。教育和研究的结合是通过两个具体的教育活动实现的，旨在通过为中学生开发互动的STEM课程和为当地女童子军实施互动示范来提高STEM的热情和意识。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.