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High level expression and characterisation of Plasmepsin II, an aspartic proteinase from Plasmodium falciparum
Author(s) -
Hill Jeffrey,
Tyas Lorraine,
Phylip Lowri H.,
Kay John,
Dunn Ben M.,
Berry Colin
Publication year - 1994
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 - 10.1016/0014-5793(94)00940-6
Subject(s) - plasmodium falciparum , biochemistry , enzyme , chemistry , protease , microbiology and biotechnology , biology , malaria , immunology
DNA encoding the last 48 residues of the propart and the whole mature sequence of Plasmepsin II was inserted into the T7 dependent vector pET 3a for expression in E. coli . The resultant product was insoluble but accumulated at ∼20 mg/l of cell culture. Following solubilisation with urea, the zymogen was refolded and, after purification by ion‐exchange chromatography, was autoactivated to generate mature Plasmepsin II. The ability of this enzyme to hydrolyse several chromogenic peptide substrates was examined; despite an overall identity of ∼35% to human renin, Plasmepsin II was not inhibited significantly by renin inhibitors.