Antonio Astarita and Francesca Muzio

Abstract:

Fluorine is rare in natural products but can be incorporated into aromatic amino acid residues of proteins via bacterial expression techniques. The incorporated fluorine can provide a useful label for NMR spectroscopy while typically not affecting the protein structure or function. We aimed to incorporate fluorine labels into a well-studied protein, glutathione S-transferase (GST). The structure and conformational changes of GST have been determined via other methods, so it can serve as a proof-of-principle for the fluorine incorporation technique. Here, we demonstrate the successful incorporation of 19F-labeled tryptophan residues into GST and optimization of expression. Future work will involve the NMR analysis of GST and the application of this technique to other proteins of interest.