Plasmodium falciparum carbonic anhydrase is a possible target for malaria chemotherapy.
Plasmodiumfalciparum is responsible for the majority of life-threatening cases of human malaria. The global emergence of drug-resistant malarial parasites necessitates identification and characterization of novel drug targets. Carbonic anhydrase (CA) is present at high levels in human red cells and in P. falciparum. Existence of at least three isozymes of the alpha < class was demonstrated in P. falciparum and a rodent malarial parasite Plasmodium berghei. The major isozyme CA1 was purified and partially characterized from P. falciparum (PfCA1). A search of the malarial genome database yielded an open reading frame similar to the alpha-CAs from various organisms, including human. The primary amino acid sequence of the PfCA1 has 60% identity with a rodent parasite Plasmodium yoelii enzyme (PyCA). The single open reading frames encoded 235 and 252 amino acid proteins for PfCA1 and PyCA, respectively. The highly conserved active site residues were also found among organisms having alpha-CAs. The PfCA1 gene was cloned, sequenced and expressed in Escherichia coli. The purified recombinant PfCA1 enzyme was catalytically active. It was sensitive to acetazolamide and sulfanilamide inhibition. Kinetic properties of the recombinant PfCA1 revealed the authenticity to the wild type enzyme purified from P. falciparum in vitro culture. Furthermore, the PfCA1 inhibitors acetazolamide and sulfanilamide showed good antimalarial effect on the in vitro growth of P. falciparum. Our molecular tools developed for the recombinant enzyme expression will be useful for developing potential antimalarials directed at P. falciparum carbonic anhydrase.
Reungprapavut
Sutarnthip
S
Unit of Biochemistry, Department of Medical Science, Faculty of Science, Rangsit University, Paholyothin Rd., Patumthani 12000, Thailand.
Krungkrai
Sudaratana R
SR
Krungkrai
Jerapan
J
D000086:Acetazolamide / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology / Q000627:therapeutic use*; D002257:Carbonic Anhydrase Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000378:metabolism / Q000469:parasitology / Q000473:pathology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D019271:Hypoxanthine / Q000378:metabolism; D007700:Kinetics; D008288:Malaria / Q000188:drug therapy* / Q000469:parasitology*; D051379:Mice; D008969:Molecular Sequence Data; D010962:Plasmodium berghei / Q000201:enzymology; D010963:Plasmodium falciparum / Q000187:drug effects / Q000201:enzymology*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment
D000962:Antimalarials; D002257:Carbonic Anhydrase Inhibitors; D011994:Recombinant Proteins; D019271:Hypoxanthine; D002256:Carbonic Anhydrases; D000086:Acetazolamide