Expression, purification, and characterization of aspartic endopeptidases: Plasmodium plasmepsins and "short" recombinant human pseudocathepsin

• Published in: Current protocols in protein science Vol. Chapter 21; p. 21.14.1
• Main Authors: Beyer, Bret B, Goldfarb, Nathan E, Dunn, Ben M
• Format: Journal Article
• Language: English
• Published: United States 01.08.2004
• Subjects: Animals; Aspartic Acid Endopeptidases - genetics; Aspartic Acid Endopeptidases - isolation & purification; Aspartic Acid Endopeptidases - metabolism; Cathepsin D - genetics; Cathepsin D - isolation & purification; Cathepsin D - metabolism; Escherichia coli - genetics; Escherichia coli - metabolism; Humans; Plasmodium - enzymology; Protein Precursors - genetics; Protein Precursors - isolation & purification; Protein Precursors - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism
• ISSN: 1934-3663
• DOI: 10.1002/0471140864.ps2114s32

The unit describes a basic protocols utilized to obtain milligram amounts of enzymatically active, pure recombinant Plasmodium plasmepsins and "short" human pseudocathepsin D. Specific details for the expression and purification of Plasmodium falciparum plasmepsin 2 and "short" human pseudocathepsin D in zymogen form are described in this chapter. The plasmepsin 2 protocols are also applicable to Plasmodium vivax, P. ovale, and P. malariae plasmepsins, as well as P. falciparum plasmepsin 4.