Expression of the essential kinase PFCDPK1 from plasmodium falciparum in toxoplasma gondii facilitates the discovery of novel antimalarial drugs

Rajshekhar Y. Gaji, Lisa Checkley, Michael L. Reese, Michael T. Ferdig, Gustavo Arrizabalaga

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We have previously shown that genetic disruption of Toxoplasma gondii calcium-dependent protein kinase 3 (TgCDPK3) affects calcium ionophore-induced egress. We examined whether Plasmodium falciparum CDPK1 (PfCDPK1), the closest homolog of TgCDPK3 in the malaria parasite P. falciparum, could complement a TgCDPK3 mutant strain. PfCDPK1 is essential and plays critical roles in merozoite development, motility, and secretion. We show that expression of PfCDPK1 in the TgCDPK3 mutant strain rescues the egress defect. This phenotypic complementation requires the localization of PfCDPK1 to the plasma membrane and kinase activity. Interestingly, PfCDPK1-expressing Toxoplasma becomes more sensitive to egress inhibition by purfalcamine, a potent inhibitor of PfCDPK1 with low activity against TgCDPK3. Based on this result, we tested eight small molecules previously determined to inhibit the kinase activity of recombinant PfCDPK1 for their abilities to inhibit ionophoreinduced egress in the PfCDPK1-expressing strain. While two of these chemicals did not inhibit egress, we found that six drugs affected this process selectively in PfCDPK1-expressing Toxoplasma. Using mutant versions of PfCDPK1 and TgCDPK3, we show that the selectivities of dasatinib and PLX-4720 are regulated by the gatekeeper residue in the ATP binding site. Importantly, we have confirmed that the three most potent inhibitors of egress in the PfCDPK1-expressing strain effectively kill P. falciparum. Thus, we have established and validated a recombinant strain of Toxoplasma that can be used as a surrogate for the discovery and analysis of PfCDPK1-specific inhibitors that can be developed as antimalarials.

Original languageEnglish (US)
Pages (from-to)2598-2607
Number of pages10
JournalAntimicrobial Agents and Chemotherapy
Volume58
Issue number5
DOIs
StatePublished - 2014

Fingerprint

Toxoplasma
Antimalarials
Plasmodium falciparum
Phosphotransferases
Merozoites
Calcium Ionophores
Falciparum Malaria
calcium-dependent protein kinase
Parasites
Adenosine Triphosphate
Binding Sites
Cell Membrane

ASJC Scopus subject areas

  • Pharmacology (medical)
  • Pharmacology
  • Infectious Diseases
  • Medicine(all)

Cite this

Expression of the essential kinase PFCDPK1 from plasmodium falciparum in toxoplasma gondii facilitates the discovery of novel antimalarial drugs. / Gaji, Rajshekhar Y.; Checkley, Lisa; Reese, Michael L.; Ferdig, Michael T.; Arrizabalaga, Gustavo.

In: Antimicrobial Agents and Chemotherapy, Vol. 58, No. 5, 2014, p. 2598-2607.

Research output: Contribution to journalArticle

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