Expression, purification, and characterization of aspartic endopeptidases: Plasmodium plasmepsins and "short" recombinant human pseudocathepsin.
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.
Beyer
Bret B
BB
University of Florida, Gainesville, Florida.
Goldfarb
Nathan E
NE
University of Florida, Gainesville, Florida.
Dunn
Ben M
BM
University of Florida, Gainesville, Florida.
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D002402:Cathepsin D / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D010961:Plasmodium / Q000201:enzymology*; D011498:Protein Precursors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D011498:Protein Precursors; D011994:Recombinant Proteins; D016282:Aspartic Acid Endopeptidases; C089643:plasmepsin; D002402:Cathepsin D