title: Characterisation of carbonic anhydrase in Plasmodium falciparum.
abstract: Here we report the existence, purification and characterisation of carbonic anhydrase in Plasmodium falciparum. The infected red cells contained carbonic anhydrase approximately 2 times higher than those of normal red cells. The three developmental forms of the asexual stages, ring, trophozoite and schizont were isolated from their host red cells and found to have stage-dependent activity of the carbonic anhydrase. The enzyme was purified to homogeneity from the crude extract of P. falciparum using multiple steps of fast liquid chromatographic techniques. It had a Mr of 32 kDa and was active in a monomeric form. The human red cell enzyme was also purified for comparison with the parasite enzyme. The parasite enzyme activity was sensitive to well-known sulfonamide-based inhibitors of both bacterial and mammalian enzymes, sulfanilamide and acetazolamide. The kinetic properties and the amino terminal sequences of the purified enzymes from the parasite and host red cell were found to be different, indicating that the purified protein most likely exhibited the P. falciparum carbonic anhydrase activity. In addition, the enzyme inhibitors had antimalarial effect against in vitro growth of P. falciparum. Moreover, the vital contribution of the carbonic anhydrase to the parasite survival makes the enzyme an attractive target for therapeutic evaluation.
authors.affiliation: Unit of Biochemistry, Department of Medical Science, Faculty of Science, Rangsit University, Paholyothin Road, 12000, Patumthani, Thailand.
authors.forename: S R
authors.initials: SR
authors.lastname: Krungkrai
authors.forename: N
authors.initials: N
authors.lastname: Suraveratum
authors.forename: S
authors.initials: S
authors.lastname: Rochanakij
authors.forename: J
authors.initials: J
authors.lastname: Krungkrai
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002256:Carbonic Anhydrases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology / Q000469:parasitology; D006801:Humans; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000201:enzymology*
chemical_list: D002256:Carbonic Anhydrases