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Development of selective inhibitor of S‐adenosylmethionine synthetase in Cryptosporidium parvum
Author(s) -
Dziobak Miranda,
Parungao Gwenn,
Viola Ronald
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.921.7
Subject(s) - antiprotozoal , cryptosporidium parvum , biology , enzyme , microbiology and biotechnology , metabolite , biosynthesis , strain (injury) , biochemistry , in vitro , anatomy
Protozoan parasites have been responsible for diseases that cause significantly high morbidity and mortality rates, as well as an economic burden. The current antiprotozoal drugs suffer from multiple liabilities related to cost, efficacy, and resistance. In this study, Cryptosporidium parvum (Cpa), the protozoan parasite that causes cryptosporidiosis – a particular concern for immunocompromised patients, was targeted for the development of an antiprotozoal drug. S‐adenosylmethionine synthetase (AdoMet synthetase ‐ metK) is the enzyme that catalyzes biosynthesis of S‐adenosylmethionine (AdoMet), an essential metabolite for all living organisms. An E. coli knockout strain (Δ metK ) used as the host for the plasmid‐containing metKCpa. Expression of this gene was shown to rescue the growth of this E. coli strain . A shorter lag phase was observed for growth in rich medium, and addition of methionine shortened the lag phase in MOPS‐glucose minimal medium. This complemented strain can now be used to screen for biologically active inhibitors against this enzyme. Furthermore, Cpa metK has been overexpressed and purified for kinetic characterization, and crystallization to study structure‐based inhibitor optimization. The results allow the further development of inhibitors for other protozoan parasites.