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High‐Throughput Virtual Screening to Identify Novel Inhibitors of 6‐Phosphogluconate Dehydrogenase in Plasmodium vivax
Author(s) -
Weber Nicole Brooke,
Trieu Carolyn,
Beckham Joshua
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.921.15
Subject(s) - plasmodium vivax , virtual screening , biology , plasmodium falciparum , computational biology , malaria , high throughput screening , sanger sequencing , drug discovery , microbiology and biotechnology , biochemistry , dna , dna sequencing , immunology
The discovery of novel drugs is warranted in the face of rising resistance developed by Plasmodium vivax parasites to a number of malaria medicines. Therefore, the principal aim of this research study is to discover unknown inhibitors for 6‐phosphogluconate dehydrogenase (Pv6PG) in the protozoan parasite P. vivax . First, the full Pv6PG coding DNA sequence was synthesized with overlapping oligo‐primers, amplified via Polymerase Chain Reaction (PCR) methods, and cloned into a pET expression vector. Following transformation into E. coli BL21(DE3), the protein was expressed and then purified with Nickel Affinity Chromatography and size exclusion chromatography. Differential Scanning Fluorimetry (DSF) assays were performed to assess the ligand‐binding interactions through shifts in the denaturation temperature. One best ranking compound from the Maybridge HitFinder library was docked using GOLD and scored 74.19, but did not exhibit a significant shift of the melting curve. To identify potential inhibitors, virtual high‐throughput screening was used. First, a homology model of the target Pv6PG was created using ICM. This docking algorithm made binding mode predictions of a ChemBridge library of over 100,000 ligands. The best ranking compounds will next be tested in wet lab using spectrophotometric assays to determine if they any have significant ability to inhibit the enzymatic functionality of Pv6PG. Successful isolation of compounds that inhibit this protein will play a role in hindering the vital biochemical pathways in the malaria‐causing organism Plasmodium vivax and could lead to greater progress in the pursuit of a novel treatment for this infectious disease. Support or Funding Information HHMI