1. Dihydrofolate Reductase (DHFR) Ruixiao ‘Ray’ Gao Department of chemistry Illinois state university 10/30/2014

2. 2 Background Found in all organisms (186 aa) Encoded by DHFR gene that exists in chromosome 5 Synthesis of purine and thymidylate Nucleic acid precursor Synthesis De novo mitochondrial thymidylate biosynthesis pathway Catalyze enzyme DNA precursor synthesis

3. DHFR in Biosynthesis Mechanism

4. 4 structure 3FRA

5. 5 Amino Acid Sequence Alignment

6. 1RX2 Secondary Structure

7. 1RX2

8. Quaternary Structure Tertiary Structure 3FL9 1RX2

9. Active Sites: Met20 Amino Acids stablize alpha helices and overall structure: Ser24 & Gly39 Gly102 & Glu112 Ser81 & Gly9 3DGA

11. 11 DHFR Mutants

12. Anticancer Properties from Antimalarial Drugs Antimalarial drugs for their anticancer potential: DHFR inhibitors resist the growth of several human cancer lines Bind with pyrimethamine and P218 Cluster analysis clustered tighly the synthetic peroxides and DHFR inhibitors. Artemisinins & paclitaxel with DHFR inhibitors kill cancer cells by inducing apoptosis

13. References Hooft, R. V. H., Guy, R. K., Chibale, K., Haynes, R. K., Peitz, I., et al. (2013) Anticancer Properties of Distinct Antimalarial Drug Classes. PLoS ONE, 8, 1-11. Agarwal, P. K., Billeter, S. R., Rajagopalan, P. T. R., Benkovic, S. J., & Schiffer, S. H. (2002) Network of coupled promoting motions in enzyme catalysis. PNAS, 99, 2794-2799. Askari, B. S., and Krajinovic, M. (2010) Dihydrofolate Reductase Gene Variations in Susceptibility to Disease and Treatment Outcomes. Current Genomics, 11, 578-583. Lyons, T. (2013) Pharmaceutical case study: Evolutionary Trace analysis of dihydrofolate reductase (DHFR). Accelrys, 1-7. Mayer, R. J., Chen, J. T., Taira, K., Fierke, C. A., & Benkovic, S. J. (1986) Importance of a hydrophobic residue in binding and catalysis by dihydrofolate reductase. Proc. Natl. Acad. Sci, 83, 7718-7720. Kwon, Y. J., Sohn, M. J., Kim, H. J., & Kim, W. G. (2014) The Lactone Form of Stachybotrydial: A New Inhibitor of Dihydrofolate Reductase from Stachybotrys sp. FN298. Biol. Pharm. Bull. 37, 1406-1410. Bradrick, T. D., Shattuck, C., Strader, M. B., Wicker, C., Eisenstein, E., and Howell, E. E. (1996) Redesigning the quaternary structure of R67 dihudrofolate reductase: Creation of an active monomer from a tetrameric protein by quadruplication of the gene. J. Biol. Chem., 271, 28031-28037. Schnell, J. R., Dyson, H. J., and Wright, P. E. (2004) Structure, Dynamics, and Catalytic Function of Dihydrofolate Reductase. Annu. Rev. Biophys. Biomol. Struct. 33, 119-40.