Spatiotemporally Resolved Proteomics through Photomediated Protein Labeling

蛋白质调节人体健康。它们介导对疾病的易感性，并帮助击退入侵的病毒和微生物。该项目旨在建立一种策略，可以测量蛋白质合成和活性随时间推移在细胞或组织中的位置而变化的变化。该方法依赖于某些蛋白质在特定条件下发光的事实。如果成功，该方法将提供新的诊断工具，并可能提供新的治疗方法。该项目还支持为高中学生编写教材。这些材料包括课堂演示和动手实验。这种经验应该介绍和吸引代表性不足的群体，以生物化学工程，并可以提高学生对科学，技术，工程和数学领域的兴趣。直接测量蛋白质身份和丰度的策略是有限的。荧光显微镜可以同时实时定位和测量细胞或组织样本中的一些蛋白质。基于质谱的蛋白质组学分析允许对数千种蛋白质进行全球测量，但只能从统一的样品中进行。所提出的策略利用非典型氨基酸，结合生物正交光化学，标记和分离蛋白质合成在指定的时间段和培养位置之前的蛋白质组学分析。分析方法能够提供任何类型的细胞或组织内蛋白质表达的定量信息。该策略使用广泛可用的仪器和开源蛋白质组学软件包，因此它们可以很容易地被世界各地的实验室采用。该奖项反映了NSF的法定使命，并被认为是值得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的支持。

Proteins regulate human health. They mediate susceptibility to illness, and they help repel invading viruses and microbes. This project seeks to establish a strategy that can measure changes to protein synthesis and activity that vary with location in the cell or tissue over time. The method depends on the fact that some proteins emit light under certain conditions. If successful, the method will provide new diagnostic tools and possibly new treatments. The project also supports the development of course materials for high school students. These materials include an in-class demonstration and a hands-on experiment. This experience should introduce and attract under-represented groups to biochemical engineering and could increase student interest in science, technology, engineering, and mathematics fields. Strategies to measure protein identity and abundance directly are limited. Simultaneous real-time location and measurement of a few proteins within cells or tissue samples is possible with fluorescence microscopy. Mass-spectrometry-based proteomic analysis permits global measurement of thousands of proteins, but only from uniform samples. The proposed strategy exploits non-canonical amino acids, in conjunction with bioorthogonal photochemistry, to tag and isolate proteins synthesized at specified periods of time and culture locations prior to proteomic analysis. The analytic methodologies are capable of providing quantitative information for protein expression within any type of cell or tissue. The strategies developed use widely available instrumentation and open-source proteomic software packages, so they could be readily adopted by laboratories worldwide.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.