Purification and characterization of Plasmodium berghei DNA topoisomerases I and II: drug action, inhibition of decatenation and relaxation, and stimulation of DNA cleavage.
It has recently been suggested that topoisomerases could be important targets for drugs used in several diseases. This prompted us to purify and characterize the topoisomerases I and II present in the erythrocytes of protozoan parasites of the genus Plasmodium, the causative agent of malaria, in order to later use these enzymatic systems in antimalarial drug assays. The topoisomerases were purified from Plasmodium berghei, a parasite of mouse red cells. The Plasmodium topoisomerase II consists of two subunits with a molecular weight of about 160K. The enzyme is ATP- and Mg2+-dependent. The conditions for the reactions of relaxation, unknotting, decatenation, and catenation were found to be similar to those observed with enzymes from other eukaryotic cells. The Plasmodium topoisomerase I is a monomeric enzyme with a Mr of 70K-100K. It is ATP-independent and K+- or Na-dependent. Mg2+ is not required for relaxation but stimulates the reaction. Topoisomerase II was more sensitive to drug action than topoisomerase I. The most active drugs were the ellipticine derivatives. The antimalarial drugs, currently used in human clinical therapy, were poor inhibitors. Some antitumoral drugs stimulated the double-stranded DNA cleavage activity of Plasmodium topoisomerase II, like that of mammalian topoisomerases II. Antimalarial drugs had no stimulating activity. It is therefore suggested that Plasmodium topoisomerases are not good targets for antimalarial drugs.
D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D004264:DNA Topoisomerases, Type I / Q000302:isolation & purification* / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D004611:Ellipticines / Q000494:pharmacology*; D005260:Female; D007700:Kinetics; D046911:Macromolecular Substances; D051379:Mice; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D010962:Plasmodium berghei / Q000201:enzymology*; D013539:Simian virus 40; D013329:Structure-Activity Relationship; D059004:Topoisomerase I Inhibitors
D000470:Alkaloids; D000962:Antimalarials; D004279:DNA, Viral; D004611:Ellipticines; D046911:Macromolecular Substances; D059004:Topoisomerase I Inhibitors; D004264:DNA Topoisomerases, Type I