Undergraduate Research at FIU (URFIU) Conference

Expression and Purification of 15N, 13Clabeled HMGA2 for NMR Structural Studies

East and Center Ballrooms

The mammalian high mobility group protein AT-hook 2 (HMGA2) is a nuclear oncoprotein, and its abnormal expression is directly linked to tumorigenesis. The HMGA2 gene is located on the chromosomal bands 12q13-15. Rearrangement of these bands disrupts the normal functions of HMGA2 and causes the formation of benign tumors, such as lipomas, and malignant tumors, such as lung cancer. In this study, our objective was to make sufficient amounts of 15N-, 13C-labeled HMGA2 for NMR studies to obtain a high-resolution structure of HMGA2-DNA complexes. In order to label this protein, E. coli strain BLR(DE3), containing a plasmid expressing HMGA2, was grown in minimal M9 medium containing 15NH4Cl and 13C-glucose. The 15N-, 13Clabeled HMGA2 protein was purified according to a purification protocol previously published by our research group. SDS-PAGE, Western Blotting, and Mass Spectrometry were used to demonstrate the successful expression and purification of the 15N-, 13C-labeled HMGA2. HMGA2 protein samples were sent to our collaborators at the University of Bayreuth, Germany for NMR structural studies.