High level expression and characterisation of Plasmepsin II, an aspartic proteinase from Plasmodium falciparum

FEBS Letters

Phylip, Lowri H.¹  Dunn, Ben M.²  Tyas, Lorraine¹  Hill, Jeffrey¹  Kay, John¹  Berry, Colin¹ 

[1] Department of Biochemistry, University of Wales College of Cardiff, PO Box 903, Cardiff CF1 1ST Wales, UK;Department of Biochemistry and Molecular Biology, J. Hillis Miller Health Center, University of Florida, Gainesville, FL 32610, USA

关键词: Aspartic proteinase;    Plasmepsin II;    Plasmodium falciparum;   

DOI  :  10.1016/0014-5793(94)00940-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown