CAREER: Optically Controlled Protein Proximity Labelling

在化学系生命过程化学项目的支持下，怀俄明州大学的Michael Taylor博士将开发新的光促进化学反应，从而能够研究线粒体转运蛋白的生物学作用。 在细胞内发现的蛋白质与邻近蛋白质形成复杂的网络，以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聚集在一起而相互作用和交流。 这些网络通常会对外部刺激做出快速反应，或者因为细胞功能障碍而发生变化，这使得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，该反应将感兴趣的蛋白质标记为与化学探针的非天然共价键。 该项目中开发的标记化学通过应用特定颜色的光来控制。 该项目通过探针的精确分子设计来利用这一要求，预计这将使迭代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的蛋白质本身，还可以研究目标蛋白质如何与相邻蛋白质网络，以及这些网络如何在真实的时间内发生变化。 综合教育目标的重点是通过两项关键活动提高怀俄明州对科学-技术-工程和数学（STEM）的认识和热情：（1）在中学一级试行在线互动课程，强调生物学和化学是如何内在联系的;以及（2）使用本项目中开发的化学反应，使用美观的荧光分子，作为当地女童子军部队互动演示的基础。 有了这个职业奖，生命过程计划的化学资助博士迈克尔·泰勒在怀俄明州大学开发新的，光促进的化学反应，应用于线粒体转运蛋白的生物学作用的研究，富含色氨酸的感觉蛋白（TSPO）。 TSPO已被证明是一种具有挑战性的蛋白质，需要通过规范的实验工作流程进行研究，因此，其在细胞功能中的作用仍然存在争议。 Taylor博士试图通过开发一种生物缀合反应来解决这一挑战，该反应结合了多种选择性因素，包括光活化，以高精度化学标记TSPO。 Taylor博士的目标是利用本文所采用的化学系统的模块化可调性来开发一套试剂，这些试剂在广泛的波长范围内被激活，并且可以通过不同的机制实现蛋白质标记。 这种新方法开辟了使用光学编排的迭代蛋白质标记序列来标记TSPO本身以及邻近蛋白质的可能性。 反过来，这可以使周围的TSPO，这将提供独特的洞察TSPO的生物作用的生物环境的三维渲染的建设。 该职业奖提供了开发，理解和应用一套化学试剂和反应的支持，这些试剂和反应允许在细胞环境中研究TSPO。 教育和研究的整合是通过两个具体的教育活动来实现的，旨在通过为中学生开发交互式STEM课程和为当地女童子军部队实施交互式演示来提高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价值和更广泛的影响审查标准进行评估而被认为值得支持。

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.