Deactivation of Protein Function and Mapping of Protein-Protein Interactions via Light-Induced Localized Oxidation

通过这一奖项，化学系的生命过程化学计划资助匹兹堡大学的Alexander Deiters教授开发新的方法来使蛋白质失活并确定蛋白质之间的相互作用。蛋白质通常由自然产生的氨基酸单位组成，在生物化学中发挥着广泛的作用，包括使化学反应更快，并与其他蛋白质相互作用，启动化学和生物事件。为了了解参与这些活动的蛋白质的具体细节，Deiters教授的团队正在开发新的光驱动化学反应，只改变涉及的关键氨基酸位点的原子，从而能够识别对化学和生物活动的成功至关重要的一个或多个蛋白质的功能。在用氨基酸中新原子的特定化学反应标记蛋白质后，通过有效地称重蛋白质，可以揭示关键的氨基酸位点。由于Deiters教授的招生策略以及所需的分子制造和蛋白质化学以及细胞生物学的结合，该项目为化学和生物学领域的下一代科学家提供了一个独特的培训环境，他们具有高度的多样性(超过60%来自代表不足的群体)。此外，为了促进公众对生物过程中光的研究和培训活动的了解和兴趣，Deiters教授和他的团队经常在当地的科学博物馆举办外展活动。这些实践推广活动包括设计实验，让孩子们在很小的时候就对科学感兴趣，同时向他们展示化学参与了一系列日常过程，比如让萤火虫发光的反应或防晒霜保护人们免受太阳有害射线的伤害。通过将光敏发色团特异性地引入靶蛋白，可以在空间和时间上产生单线态氧，从而能够高精度地可靠地改变蛋白质的结构和功能。该方法是基于非天然氨基酸的突变，然后是生色团生物结合，从而提供对单线态产氧生色团的放置的完全可编程控制。这提供了对蛋白质表面景观和活性部位内产生的活性氧物种的接近程度的精细控制。除了光触发的蛋白质功能失活，这项技术实现的定向蛋白质氧化还允许精确的基于邻近的标记和下拉，导致识别和绘制蛋白质-蛋白质相互作用的地图。这一奖项反映了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 this award, the Chemistry of Life Processes Program in the Division of Chemistry is funding Professor Alexander Deiters at the University of Pittsburgh to develop new approaches to deactivate proteins and identify protein-protein interactions. Proteins, which are typically made up of naturally occurring amino acid units, play a wide range of roles in biological chemistry, including making chemical reactions faster and interacting with other proteins to start chemical and biological events. To understand the specific details of proteins involved in these events, Professor Deiters' team is developing new light-driven chemical reactions that change the atoms of only the key amino acid sites involved, which allows identification of the function of the protein or proteins crucial to the success of the chemical and biological events. The key amino acid sites are revealed by effectively weighing proteins after they have been marked with chemical reactions specific to the new atoms in the amino acids. Due to Professor Deiters' student recruiting strategies and the required combination of molecule making and protein chemistry, and cell biology, this project provides a unique training environment for the next generation of scientists at the interface of chemistry and biology, with them being highly diverse (greater than 60% from under-represented groups). Furthermore, to foster public understanding and interest in the research and training activities focused on light in biological processes, Professor Deiters and his team routinely offer outreach activities at local science museums. The hands-on outreach activities include the design of experiments that excite children at an early age about science, while showing them the involvement of chemistry in a range of everyday processes, such as reactions that make fireflies glow or sunscreens protect people from the sun's harmful rays. The site-specific introduction of photosensitive chromophores into target proteins enables spatial and temporal generation of singlet oxygen capable of reliably modifying protein structure and function with high precision. The approach is based on unnatural amino acid mutagenesis, followed by chromophore bioconjugation, thereby providing fully programmable control over the placement of singlet oxygen-generating chromophores. This affords exquisite control over the proximity of reactive oxygen species produced within protein surface landscapes and active sites. In addition to the light-triggered deactivation of protein function, the targeted protein oxidation enabled by this technology also allows for precise proximity-based labeling and pull-down, leading to the identification and mapping of protein-protein interactions.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.