Monica Eldeiry, Sara Thaqi and Naveen Mufti

Abstract:

Fluorine, uncommon in biological molecules, can be incorporated into aromatic amino acid residues of proteins using bacterial expression techniques. The fluorine provides a sensitive NMR probe to investigate protein structure and conformation without disrupting protein activity. Our group aims to use this methodology to study conformational changes of the histone methyltransferase PRDM2, a cancer-associated protein involved in cell division and differentiation. Prior studies of similar proteins suggest that conformational changes of PRDM2 are necessary for catalytic activity, but available X-ray and solution structures lack the flexible region of the catalytic domain. We hope to use fluorine-19 NMR spectroscopy to provide structural evidence for the proposed conformational changes in wild type PRDM2. Using minimal media supplemented with 4-, 5-, or 6-fluoroindole, we optimized small-scale expression of the catalytic domain of His-tagged PRDM2 and two enzymatically deficient variants found previously in cancer cell lines. We also began work on large-scale expression and purification of fluorine-labeled PRDM2 to provide protein for future NMR analysis.