STRUCTURAL STUDY OF PRDM2:
PR domain-containing (PRDM) proteins, a large sub-class of HKMTs, are important in cell division and differentiation. Aberrant methylation has been demonstrated in various disease states; therefore, knowledge of the structure and kinetics of the enzymes can aid in understanding the disease mechanism. Work on a similar enzyme showed that a conformational change in the enzyme is needed to create the active site and enable methyltransferase activity. While NMR and X-ray crystal structures of PRDM2 have been solved, all structures involved inactive truncations of the enzyme that had cleaved the amino acids likely to be involved in the conformational change. The goal of this work is to assess structural changes in kinetically active wild type and mutant variants of PRDM2 upon addition of substrates via NMR, FT-IR, and nanoDSC. This research will not only provide the students in our group an opportunity to learn a variety of scientific techniques but will also engage them in a novel area of research relevant to human health and disease.
Structural study of known PRDM2 variants found in human cancer:
PRDM2 missense mutations have been found in human cancers. Three of these (I188V, A159V, and C106Y) are located in or directly adjacent to the methyltransferase domain of the enzyme. While the net negative effect of the point mutations toward HKMT activity has been demonstrated, there has been no investigation into the biochemical mechanism behind the decease in H3 methylation. Our group is expressing and purifying the catalytic domain of the known variants of PRDM2 found in the cancer cells. The domain structures with and without substrates will be studied for helical content and thermal stability using FT-IR, nanoDSC, and NMR and compared to the wild-type PRDM2 methyltransferase domain.