Utility of (His)6 tag for purification and refolding of proplasmepsin-2 and mutants with altered activation properties

Protein Expression and Purification
S V GulnikJohn W Erickson

Abstract

Plasmepsin-2 is a malarial aspartic proteinase that has been implicated in the initial steps of hemoglobin degradation in parasites and thus represents an attractive antimalarial target. Escherichia coli expressed proplasmepsin-2 is capable of activation at acidic pH by autocatalytic cleavage of the pro part region, which results in products of different length. We designed a 10-amino-acid deletion in the pro part region that allows faster generation of homogeneous enzyme upon activation. Incorporation of a (His)6 tag onto the N-terminus of the pro part enables on-column refolding of proplasmepsin-2 and simplifies proenzyme purification and pro part separation after activation. The proposed purification procedure results in highly pure and easily crystallizable enzyme.

References

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Citations

Jul 11, 2009·Protein Expression and Purification·Ariun Narmandakh, Stephen L Bearne
Dec 2, 2015·Acta Crystallographica. Section F, Structural Biology Communications·Rosario RecachaKaspars Tars
Jan 12, 2005·Protein Expression and Purification·Jin-Cun ZhaoXiao-Ming Gao
Jun 3, 2006·Current Opinion in Biotechnology·Wieslaw Swietnicki
Jun 6, 2014·ACS Medicinal Chemistry Letters·Kristaps JaudzemsAigars Jirgensons
Apr 23, 2008·Current Protocols in Protein Science·Paul T Wingfield
Apr 2, 2015·Current Protocols in Protein Science·Paul T Wingfield

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