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.
