title: Functional expression of falcipain, a Plasmodium falciparum cysteine proteinase, supports its role as a malarial hemoglobinase.
abstract: Erythrocytic malaria parasites degrade hemoglobin as a principal source of amino acids for parasite protein synthesis. We have previously shown that a Plasmodium falciparum trophozoite cysteine proteinase, now termed falcipain, is required for hemoglobin degradation, and we have hypothesized that this proteinase is responsible for initial cleavages of hemoglobin. To further evaluate the biological role of falcipain, we expressed the enzyme in bacterial and viral expression systems. After expression in the baculovirus system, falcipain was enzymatically active and had biochemical properties very similar to those of the native proteinase. Recombinant falcipain rapidly hydrolyzed both denatured and native hemoglobin. Hemoglobin hydrolysis was blocked by cysteine proteinase inhibitors but not by inhibitors of other classes of proteinases. Our results support our hypothesis that falcipain is a critical malarial hemoglobinase that is responsible for both initial cleavages of hemoglobin and the subsequent hydrolysis of globin into small peptides.
authors.affiliation: Department of Medicine, San Francisco General Hospital, University of California 94143, USA.
authors.forename: F
authors.initials: F
authors.lastname: Salas
authors.forename: J
authors.initials: J
authors.lastname: Fichmann
authors.forename: G K
authors.initials: GK
authors.lastname: Lee
authors.forename: M D
authors.initials: MD
authors.lastname: Scott
authors.forename: P J
authors.initials: PJ
authors.lastname: Rosenthal
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics; D006454:Hemoglobins / Q000378:metabolism; D010963:Plasmodium falciparum / Q000201:enzymology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis
chemical_list: D006454:Hemoglobins; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases; C099607:falcipain