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

通过光诱导局部氧化使蛋白质功能失活并绘制蛋白质-蛋白质相互作用图

• 批准号: 1904972
• 负责人: Alexander Deiters
• 金额: $ 42万
• 依托单位: University of Pittsburgh
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1904972&HistoricalAwards=false
• 关键词: Deactivation; Protein; Function; Mapping; Interactions

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.

有了这个奖项，化学系的生命过程化学计划正在资助匹兹堡大学的亚历山大迪特斯教授开发新的方法来灭活蛋白质和识别蛋白质-蛋白质相互作用。蛋白质通常由天然存在的氨基酸单元组成，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以启动化学和生物事件。为了了解参与这些事件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，这些反应仅改变所涉及的关键氨基酸位点的原子，这使得能够识别蛋白质的功能或对化学和生物事件的成功至关重要的蛋白质。关键的氨基酸位点是通过对蛋白质进行有效的称重来揭示的，蛋白质是用氨基酸中新原子特有的化学反应来标记的。由于Deiters教授的学生招募策略以及分子制造和蛋白质化学以及细胞生物学的必要组合，该项目为化学和生物学界面的下一代科学家提供了独特的培训环境，他们高度多样化（超过60%来自代表性不足的群体）。此外，为了促进公众对以生物过程中的光为重点的研究和培训活动的理解和兴趣，Deiters教授及其团队定期在当地科学博物馆举办外展活动。实践推广活动包括设计实验，激发儿童在早期对科学的兴趣，同时向他们展示化学在一系列日常过程中的参与，例如使萤火虫发光或防晒霜保护人们免受太阳有害射线的反应。将光敏发色团位点特异性地引入靶蛋白质中使得能够在空间和时间上产生单线态氧，从而能够以高精度可靠地修饰蛋白质结构和功能。该方法是基于非天然氨基酸诱变，然后通过发色团生物共轭，从而提供完全可编程控制的单线态氧产生发色团的位置。这提供了对蛋白质表面景观和活性位点内产生的活性氧物种的接近度的精细控制。除了光引发的蛋白质功能失活，该技术实现的靶向蛋白质氧化还允许精确的基于邻近的标记和下拉，从而识别和绘制蛋白质-蛋白质相互作用。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Targeted protein degradation through light-activated E3 ligase recruitment

通过光激活 E3 连接酶募集进行靶向蛋白质降解

• 发表时间: 2023
• 期刊: Methods in enzymology
• 作者: Olivia Shade, Amy Ryan

Quantitative Analysis and Optimization of Site-Specific Protein Bioconjugation in Mammalian Cells.

• DOI: 10.1021/acs.bioconjchem.2c00451
• 发表时间: 2022-12
• 期刊: Bioconjugate chemistry
• 影响因子: 4.7
• 作者: Amy Ryan;Olivia Shade;Anirban Bardhan;Aleksander Bartnik;A. Deiters

Targeted Protein Degradation through Fast Optogenetic Activation and Its Application to the Control of Cell Signaling

• DOI: 10.1021/jacs.1c04324
• 发表时间: 2021-06-14
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Ryan, Amy;Liu, Jihe;Deiters, Alexander
• 通讯作者: Deiters, Alexander