Plasmodium falciparum Purine Nucleoside Phosphorylase Is Critical for Viability of Malaria Parasites
| Autor: | Vern L. Schramm, Dennis C Madrid, Karena L. Waller, Li-Min Ting, Kami Kim |
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| Rok vydání: | 2008 |
| Předmět: |
Pyrrolidines
Plasmodium falciparum Protozoan Proteins Purine nucleoside phosphorylase Pyrimidinones Biochemistry Animals Genetically Modified chemistry.chemical_compound Adenosine deaminase Transition state analog Animals Humans Enzyme Inhibitors Malaria Falciparum Purine metabolism Molecular Biology Nucleotide salvage Hypoxanthine biology Transition (genetics) Cell Biology biology.organism_classification Metabolism and Bioenergetics Purine-Nucleoside Phosphorylase chemistry Purines Gene Knockdown Techniques biology.protein |
| Zdroj: | Journal of Biological Chemistry. 283:35899-35907 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m807218200 |
| Popis: | Human malaria infections resulting from Plasmodium falciparum have become increasingly difficult to treat due to the emergence of drug-resistant parasites. The P. falciparum purine salvage enzyme purine nucleoside phosphorylase (PfPNP) is a potential drug target. Previous studies, in which PfPNP was targeted by transition state analogue inhibitors, found that those inhibiting human PNP and PfPNPs killed P. falciparum in vitro. However, many drugs have off-target interactions, and genetic evidence is required to demonstrate single target action for this class of potential drugs. We used targeted gene disruption in P. falciparum strain 3D7 to ablate PNP expression, yielding transgenic 3D7 parasites (Δpfpnp). Lysates of the Δpfpnp parasites showed no PNP activity, but activity of another purine salvage enzyme, adenosine deaminase (PfADA), was normal. When compared with wild-type 3D7, the Δpfpnp parasites showed a greater requirement for exogenous purines and a severe growth defect at physiological concentrations of hypoxanthine. Drug assays using immucillins, specific transition state inhibitors of PNP, were performed on wild-type and Δpfpnp parasites. The Δpfpnp parasites were more sensitive to PNP inhibitors that bound hPNP tighter and less sensitive to MT-ImmH, an inhibitor with 100-fold preference for PfPNP over hPNP. The results demonstrate the importance of purine salvage in P. falciparum and validate PfPNP as the target of immucillins. |
| Databáze: | OpenAIRE |
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