Virtual Screening for novel inhibitors of Plasmodium vivax 6-phosphogluconate dehydrogenase

• In Event: Poster Presentation and Judging for: Biomedical; Cell & Molecular Biology; Chemistry & Biochemistry; Marine Science; Systematics & Evolutionary Biology.
  In Poster Session: Biomedical Sciences Poster Session

Abstract

6-Phosphogluconate dehydrogenase (6PGD) is a vital enzyme in the pentose phosphate pathway that converts 6-phosphogluconate to ribulose-5-phosphate. It can also serve as a potential drug target for Plasmodium vivax, a prominent malaria parasite that is transmitted by the Anopheles mosquito, infecting 19-50 million people in developing countries each year. Targeting the activity of this enzyme may decrease the survival of this protozoan parasite by inhibiting its ability to synthesize nucleotides and produce energy via glycolysis. To find a potential therapeutic molecule, virtual drug screening was used as a cost-effective method to make predictions about which compounds have the best binding affinity for the active site of 6PGD. A control set of compounds was used to validate the molecular docking software for this enzyme. Soon after, large libraries of over 25,000 compounds were screened and the top-scoring compounds were selected then filtered by Lipinski’s Rule of 5. To make protein for the validation of these top compounds in inhibition assays, Plasmodium vivax 6PGD coding DNA sequence was cloned into the pNIC-Bsa4 expression vector, expressed in BL21(DE3) Competent E.coli cells, and purified using His Tag Protein Purification (Ni-NTA) and Anion Exchange Chromatography. Differential Scanning Fluorimetry (DSF) was used to determine the melting point of the enzyme and the binding affinity of the ligand to the active site. Enzyme and inhibition assays of 6PGD were also done with the selected compounds and the purified protein to determine if the compounds changed the enzyme activity of 6PGD.

Authors

• Geovanny Antonio Zarceno, 1997

• Josh Beckham, Research Educator

• Walter Fast, Principal Investigator