Characterisation of carbonic anhydrase in Plasmodium falciparum

Malaria parasite carbonic anhydrase: inhibition of aromatic/heterocyclic sulfonamides and its therapeutic potential

“…The recombinant pfCA protein shows authenticity to the native enzyme purified from in vitro P. falciparum culture [28] . The kinetic parameters including KM, KCAT, KI of the inhibitor AZA are also found to be similar between the native and recombinant enzymes ( Table 2).…”

“…CA was first purified from bovine red cells in 1933 [12] , followed by the identification of several isozymes ubiquitously distributed in all organisms so far examined [13][14][15][16][17][18][19] . The CAs in protozoa and helminthes parasites are found sparsely [24][25][26][27][28] . Recent studies on the biochemistry and the crystal structure of CAs from various organisms reveal that they evolved independently al least five times, with five genetically distinct enzyme families known to date: the 毩 -, 毬 -, 毭 -, 毮 -, and 毱 -CAs [29][30][31][32] .…”

“…During 2001-2004, we demonstrated the existence of the CA enzymatic activities in P. falciparum from in vitro culture and in the mouse parasite P. berghei [28,59].…”

“…The most effective CAIs for pfCA were also assayed in vitro, for the inhibition of the parasite growth in cell cultures, as well as in vivo, in an animal model of drug testing for human malaria, i.e., mice infected with P. berghei. Table 1 Stage-dependent 毩 -carbonic anhydrase activity in three developmental forms of P. falciparum, comparing to normal and infected red cells [28] . AZA and SFA are acetazolamide and sulfanilamide (4-aminobenzenesulfonamide), respectively [59] .…”

A magnificent enzyme superfamily: carbonic anhydrases, their purification and characterization

“…In this procedure only the hCAII molecules that possess activity bound to the column, whereas aggregated and misfolded proteins washed through the column 62 . Krungkrai et al applied a three steps purification procedure of the malarial CA (pfCA) by using combinations of cationexchange Mono S column (fractions eluting at pH 6.5) and anion-exchange Mono Q column (fractions eluting at 80 mM KCl), as well as gel filtration on Superose 12 column or on a FPLC system 63 . Different from other studies, cytosolic and integral CA isozymes were purified with a Sepharose-4B-L-TyrosineSulfanilamide affinity column and characterized extensively 64 .…”