Synthesis and Characterization of Non-Natural Fluorescent Amino Acids
Arts and Sciences
VU Chemistry Department
In the past decade, fluorescent proteins (FPs), such as GFP, have revolutionized the study of microscopic and nanoscopic biology. The past twenty years has focused on producing many derivatives and analogs of FPs that have allowed biologists to enter into the cellular world. Similar work has been completed on the synthesis of non-natural fluorescent amino acids (NFAAs). A long term goal of our research is to create a cellular screening mechanism that could use tRNA/aaRS pairs that could incorporate NFAAs into CooA that could serve as a carbon monoxide detector in neurobiological environments. CooA is a heme-containing carbon monoxide (CO) sensor from the bacterium Rhodospirillum rubrum. Carbon monoxide has long been known to be an important neurotransmitter in brain cells. When CooA binds carbon monoxide, it undergoes a conformational shift. If a NFAA were incorporated into CooA via a cellular screening mechanism, the protein will could expose NFAA to the cellular environment and fluoresce. Thus, carbon monoxide activity could be visualized in vivo instead of in vitro. The focus of this paper’s research is the synthesis and characterization of L-alanine,3-[7-nitro-2,1,3-benzoxadiazol-4-yl], a NFAA analog of tryptophan. The synthesis followed Katritzky and Narindoshvilli’s 2009 procedure. After an initial round of synthesis and characterization, we found results that suggested our synthesis was successful and that L-alanine,3-[7-nitro-2,1,3-benzoxadiazol-4-yl] was present in the product solution. Future work requires further analysis and characterization, as well as repeated syntheses.
Hoemann, Benjamin L., "Synthesis and Characterization of Non-Natural Fluorescent Amino Acids" (2015). Symposium on Undergraduate Research and Creative Expression (SOURCE). 447.