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Recombinant Plasmodium falciparum glutathione reductase is inhibited by the antimalarial dye methylene blue
Author(s) -
Färber P.M,
Arscott L.D,
Williams C.H,
Becker K,
Schirmer R.H
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00031-3
Subject(s) - methylene blue , plasmodium falciparum , recombinant dna , glutathione , chemistry , enzyme , escherichia coli , biochemistry , biology , malaria , photocatalysis , gene , immunology , catalysis
Plasmodium falciparum glutathione reductase (PfGR) has emerged as a drug target against tropical malaria. Here we report the expression of PfGR in Escherichia coli SG5(DE3) and isolation procedures for this protein. Recombinant PfGR does not differ from the authentic enzyme in its enzymic properties, the turnover number being 9900 min −1 . The dimeric flavoenzyme exhibits redox‐dependent absorption spectra; the single tryptophan residue (per 57.2 kDa subunit) is strongly fluorescent. PfGR can be inhibited by the antimalarial drug methylene blue at therapeutic concentrations; the K i for non‐competitive inhibition is 6.4 μM. The sensitivity to methylene blue is observed also at high ionic strength so that, by analogy to human GR, analysis of crystalline enzyme‐drug complexes can be envisaged.