title: dUTPase as a platform for antimalarial drug design: structural basis for the selectivity of a class of nucleoside inhibitors.
abstract: Pyrimidine metabolism is a major route for therapeutic intervention against malaria. Here we report inhibition and structural studies on the deoxyuridine nucleotidohydrolase from the malaria parasite Plasmodium falciparum (PfdUTPase). We have identified a series of triphenylmethane derivatives of deoxyuridine with antimalarial activity in vitro which inhibit specifically the Plasmodium dUTPase versus the human enzyme. A 2.4 Angstrom crystal structure of PfdUTPase in complex with one of these inhibitors reveals an atypical trimeric enzyme in which the triphenylmethane derivative can be seen to select for PfdUTPase by way of interactions between the trityl group and the side chains of residues Phe46 and Ile117. Immunofluorescence microscopy studies of parasitized red blood cells reveal that enzyme concentrations are highest during the trophozoite/schizont stages, suggesting that PfdUTPase has a major role in DNA replication. Taken together the data show that PfdUTPase may be considered as an antimalarial drug target.
authors.affiliation: Structural Biology Laboratory, Department of Chemistry, University of York, Heslington, York YO10 5YW, United Kingdom.
authors.forename: Jean L
authors.initials: JL
authors.lastname: Whittingham
authors.forename: Isabel
authors.initials: I
authors.lastname: Leal
authors.forename: Corinne
authors.initials: C
authors.lastname: Nguyen
authors.forename: Ganasan
authors.initials: G
authors.lastname: Kasinathan
authors.forename: Emma
authors.initials: E
authors.lastname: Bell
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Jones
authors.forename: Colin
authors.initials: C
authors.lastname: Berry
authors.forename: Agustin
authors.initials: A
authors.lastname: Benito
authors.forename: Johan P
authors.initials: JP
authors.lastname: Turkenburg
authors.forename: Eleanor J
authors.initials: EJ
authors.lastname: Dodson
authors.forename: Luis M
authors.initials: LM
authors.lastname: Ruiz Perez
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Wilkinson
authors.forename: Nils Gunnar
authors.initials: NG
authors.lastname: Johansson
authors.forename: Reto
authors.initials: R
authors.lastname: Brun
authors.forename: Ian H
authors.initials: IH
authors.lastname: Gilbert
authors.forename: Dolores
authors.initials: D
authors.lastname: Gonzalez Pacanowska
authors.forename: Keith S
authors.initials: KS
authors.lastname: Wilson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000962:Antimalarials / Q000737:chemistry* / Q000494:pharmacology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000201:enzymology*; D017433:Protein Structure, Secondary; D011755:Pyrophosphatases / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D014320:Triphenylmethyl Compounds / Q000737:chemistry
chemical_list: D000962:Antimalarials; D004791:Enzyme Inhibitors; D014320:Triphenylmethyl Compounds; C046945:triphenylmethane; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase