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Expression, Purification, And Characterization of W. Bancrofti DHFR and its Evaluation as a Possible Drug Target for Treating Lymphatic Filariasis
Author(s) -
Tobias Andrew M,
Gubler Ulrich A,
Goodey Nina
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.921.2
Subject(s) - lymphatic filariasis , wuchereria bancrofti , mass drug administration , lymphatic system , biology , tropical disease , enzyme , filariasis , parasitic disease , neglected tropical diseases , disease , immunology , biochemistry , medicine , helminths , environmental health , population
Lymphatic filariasis is a disease caused by filarial nematodes that affects millions of individuals every year. A specific filarial nematode, W. bancrofti, is one organism that causes Lymphatic filariasis. This disease affects the lymphatic system and can cause severe swelling in the lower appendages. According to the World Health Organization, people living in Bangladesh, India, Indonesia, Myanmar, Nigeria, Nepal, the Philippines and others are at risk of infection by this neglected tropical disease. To help cure those infected, research is needed to find a drug able to inhibit the proliferation of W. bancrofti . The protein we are evaluating in order to possibly accomplish this task is WbDHFR. DHFR is an enzyme that reduces dihydrofolate into tetrahydrofolate (THF). THF and its derivatives are required for purine synthesis, making the enzyme crucial for cell proliferation. In this project, WbDHFR was expressed in LOBSTR strain E. coli and purified using a Hexa‐His tag and Ni‐NTA resin. The purified enzyme is active with a V max of 159 ± 9 nM/s and K m of 36 ± 4. The enzyme will be tested against several antifolate compounds as inhibitors. Support or Funding Information Science Honors Innovation Program at Montclair State University, and the American Society for Biochemistry and Molecular Biology