Characterization of Plasmodium falciparum Calcium-dependent Protein Kinase 1 (PfCDPK1) and Its Role in Microneme Secretion during Erythrocyte Invasion

Bansal, Abhisheka; Singh, Shailja; More, Kunal R.; Hans, Dhiraj; Nangalia, Kuldeep; Yogavel, Manickam; Sharma, Amit; Chitnis, Chetan E.

doi:10.1074/jbc.m112.411934

Cited by 88 publications

(103 citation statements)

References 30 publications

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“…Our finding that staurosporine inhibits PfCDPK1 function within T. gondii at concentrations that do not affect wild-type parasites suggests that PfCDPK1 might be one of the major targets of this drug in Plasmodium. In this respect, the effects of staurosporine on P. falciparum mentioned above are consistent with the roles of PfCDPK1 in microneme secretion, motility, invasion, and intracellular development (13,17). As evident in our experiments using zaprinast, however, staurosporine might target other kinases, including PKG.…”

Section: Discussionsupporting

confidence: 71%

“…Although some of the sites phosphorylated in vitro by PfCDPK1 are also modified in vivo (18), these phosphorylated sites appear to be irrelevant to in vivo functions (44). Inhibition of PfCDPK1 has been reported to inhibit microneme secretion and invasion (17,18). On the other hand, TgCDPK3-deficient T. gondii does not exhibit major defects in either motility or microneme secretion, suggesting that TgCDPK3 substrates are likely different from those of PfCDPK1 in Plasmodium (22).…”

Section: Discussionmentioning

confidence: 90%

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Expression of the Essential Kinase PfCDPK1 from Plasmodium falciparum in Toxoplasma gondii Facilitates the Discovery of Novel Antimalarial Drugs

Gaji

Checkley

Reese

et al. 2014

Antimicrob Agents Chemother

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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.

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Section: Discussionsupporting

confidence: 71%

Section: Discussionmentioning

confidence: 90%

Expression of the Essential Kinase PfCDPK1 from Plasmodium falciparum in Toxoplasma gondii Facilitates the Discovery of Novel Antimalarial Drugs

Gaji

Checkley

Reese

et al. 2014

Antimicrob Agents Chemother

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“…CDPKs have been implicated in a wide range of functions including invasion, gametogenesis, gliding motility, egress and microneme secretion [10,11,[39][40][41]. Although a number of small molecule screens have identified pharmacological inhibitors against members of this kinase family [10,42,43], chemical genetics has played a prominent role in providing insights into the functional role of the CDPKs ( Table 1).…”

Section: Chemical Genetic Approach To the Study Of Calcium-dependent mentioning

confidence: 99%

“…ACCEPTED MANUSCRIPT 11 combining compounds 1 or 2 with wild type and mutant parasites it was possible to dissect the onand off-target actions of the inhibitors and thereby precisely define the cellular role of PKG [7,35].…”

Section: Accepted Manuscriptmentioning

confidence: 99%

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Applying chemical genetic tools to the study of phospho-signalling pathways in malaria parasites

Mitcheson

Tobin

Alam

2015

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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TargetingPlasmodiumwith constrained peptides and peptidomimetics

Helton

Kennedy

2020

IUBMB Life

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Malaria remains a worldwide health concern with an estimated quarter of a billion people infected and nearly half a million deaths annually. Malaria is caused by a parasite infection from Plasmodium strains which are transmitted from mosquitoes into the human host. Although several small molecule inhibitors have been found to target the early stages of transmission and prevent parasite proliferation, multiple drug resistant parasite strains have emerged and drug resistance remains a major hurdle. As an alternative to small molecule inhibition, several peptide-based therapeutics have been explored for their potential as antimalarial compounds. Chemically constrained peptides or peptidomimetics were developed to target large binding interfaces of parasite-based proteins that have historically been difficult to selectively inhibit using small molecules. Here, we review ongoing research aimed at developing constrained peptides targeting protein-protein interactions pertinent to malaria pathogenesis. These targets include Falcipain-2, the J domain of CDPK1, myosin A tail domain interacting protein, the PKA signaling pathway, and an unclear signaling pathway involving angiotensin-derived peptides. Diverse synthetic methods were also used for each target. Merging parasite biology with synthetic strategies may provide new opportunities to develop alternative methods for uncovering novel antimalarials and may offer an alternate source for targeting drug-resistant parasite strains.

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Characterization of Plasmodium falciparum Calcium-dependent Protein Kinase 1 (PfCDPK1) and Its Role in Microneme Secretion during Erythrocyte Invasion

Cited by 88 publications

References 30 publications

Expression of the Essential Kinase PfCDPK1 from Plasmodium falciparum in Toxoplasma gondii Facilitates the Discovery of Novel Antimalarial Drugs

Expression of the Essential Kinase PfCDPK1 from Plasmodium falciparum in Toxoplasma gondii Facilitates the Discovery of Novel Antimalarial Drugs

Applying chemical genetic tools to the study of phospho-signalling pathways in malaria parasites

TargetingPlasmodiumwith constrained peptides and peptidomimetics

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