Purification of human malaria parasite hypoxanthine guanine xanthine phosphoribosyltransferase (HGXPRT) using immobilized Reactive Red 120.
Malaria is caused by Plasmodium parasite infection. The human malarial parasite does not have a de novo pathway for synthesis of nucleotides and the purine salvage pathway enzyme hypoxanthine guanine xanthine phosphoribosyltransferase (HGXPRT) is critical for survival. In our efforts to find inhibitors of the malarial parasite HGXPRT, we have developed a simple but effective purification protocol for this protein expressed in Escherichia coli without an affinity tag. The protocol consists of tandem columns of anion exchange and immobilized Reactive Red 120 resins. The enzyme is inactive as isolated but can be activated by incubation with substrate(s).
Mbewe
Boniface
B
Chemistry Department, School of Natural Sciences, University of Zambia, P.O. Box 32379 , Lusaka, Zambia. bmbewe@gmail.com
Chibale
Kelly
K
McIntosh
David B
DB
D000818:Animals; D004396:Coloring Agents; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007700:Kinetics; D008288:Malaria; D010430:Pentosyltransferases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010961:Plasmodium / Q000201:enzymology*; D015800:Protozoan Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014227:Triazines
D004396:Coloring Agents; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D014227:Triazines; C042516:reactive red 120 dye; D010430:Pentosyltransferases; C083867:hypoxanthine-guanine-xanthine phosphoribosyltransferase