Purification and characterization of DNA polymerases from Plasmodium falciparum.
Fractionation of Plasmodium falciparum cellular extracts by fast protein liquid chromatography (FPLC) identified at least two different DNA polymerases. An aphidicolin-sensitive activity co-purified with a primase activity. This, in combination with other characteristics (processivity, sensitivity to other inhibitors), most likely classifies this enzyme as an alpha-like DNA polymerase. It was, however, relatively resistant to N2-(p-n-butylphenyl)deoxyguanosine 5'-triphosphate (IC50 = 6.6 microM) and differs in this aspect from the host homologue, possibly indicating structural differences between host and parasite DNA polymerase alpha. The other DNA polymerase matched eukaryotic DNA polymerase gamma in all properties tested.
Chavalitshewinkoon
P
P
Department of Parasitology and Tropical Veterinary Medicine, University of Utrecht, The Netherlands.
de Vries
E
E
Stam
J G
JG
Franssen
F F
FF
van der Vliet
P C
PC
Overdulve
J P
JP
D000818:Animals; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D019915:DNA Primase; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism*; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000469:parasitology; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D010963:Plasmodium falciparum / Q000201:enzymology*; D012316:RNA Nucleotidyltransferases / Q000378:metabolism
D017886:Durapatite; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004259:DNA-Directed DNA Polymerase