Molecular Characterization and Mutational Analysis of Recombinant Diadenosine 5′,5″-P&lt;sup&gt;1&lt;/sup&gt;,P&lt;sup&gt;4&lt;/sup&gt;-Tetraphosphate Hydrolase from &lt;i&gt;Plasmodium falciparum&lt;/i&gt;

Osman, Waleed; Endo, Shunsuke; Oh‐hashi, Kentaro; Kitamura, Yoshiaki; Kitade, Yukio

doi:10.1248/bpb.b12-00165

Cited by 6 publications

(11 citation statements)

References 18 publications

(16 reference statements)

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“…We performed Pf Ap4AH enzyme assays to access activity of recombinant enzyme ( Fig. 3C ), which displayed kinetic parameters similar to the earlier reports (data not shown) 11 . Enzyme assays in the presence of suramin suggested inhibition with an IC 50 value of ~11.8 μM ( Fig.…”

Section: Resultssupporting

confidence: 67%

“…Based on sequence analysis the P. falciparum Ap4AH ( Pf Ap4AH, EC 3.6.1.17 ) was predicted to be an animal-archeal type which cleaves polyphosphate chain at the fourth phosphate from the tightly bound adenosine resulting in asymmetrical cleavage of Ap4A. This is distinct from some plant-bacterial type hydrolases 8 9 11 . Eukaryotic Ap4AHs are predominantly cytoplasmic or nuclear, while the bacterial Ap4AHs appear to be ribosome associated 12 13 14 .…”

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confidence: 99%

“…It was recently shown that Ap4A can be produced in P. falciparum by lysine-tRNA synthetase ( Pf KRS) 23 . Also, the enzyme kinetics and reaction specificities of Pf Ap4AH have been studied previously 11 . Pf Ap4AH can asymmetrically hydrolase Ap4A and Ap5A molecules produced by Pf KRS (and probably by other Pf aaRSs) to ATP + AMP and ATP + ADP, respectively 8 10 .…”

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confidence: 99%

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Structural and functional attributes of malaria parasite diadenosine tetraphosphate hydrolase

Sharma

Yogavel

2016

Sci Rep

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Malaria symptoms are driven by periodic multiplication cycles of Plasmodium parasites in human red blood corpuscles (RBCs). Malaria infection still accounts for ~600,000 annual deaths, and hence discovery of both new drug targets and drugs remains vital. In the present study, we have investigated the malaria parasite enzyme diadenosine tetraphosphate (Ap4A) hydrolase that regulates levels of signalling molecules like Ap4A by hydrolyzing them to ATP and AMP. We have tracked the spatial distribution of parasitic Ap4A hydrolase in infected RBCs, and reveal its unusual localization on the infected RBC membrane in subpopulation of infected cells. Interestingly, enzyme activity assays reveal an interaction between Ap4A hydrolase and the parasite growth inhibitor suramin. We also present a high resolution crystal structure of Ap4A hydrolase in apo- and sulphate- bound state, where the sulphate resides in the enzyme active site by mimicking the phosphate of substrates like Ap4A. The unexpected infected erythrocyte localization of the parasitic Ap4A hydrolase hints at a possible role of this enzyme in purinerigic signaling. In addition, atomic structure of Ap4A hydrolase provides insights for selective drug targeting.

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Section: Resultssupporting

confidence: 67%

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confidence: 99%

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confidence: 99%

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Structural and functional attributes of malaria parasite diadenosine tetraphosphate hydrolase

Sharma

Yogavel

2016

Sci Rep

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“…[6] In order to verify ac onservede nzymatic activity for PfA-p3Aase as an A3pA hydrolase and to establisha ne xperimental system for validation of cyclomarin Aa sa ni nhibitor of the enzymatica ctivity of PfAp3Aase in vitro, we expressed three diadenosine polyphosphate hydrolases in E. coli and established enzymatic assays with use of al uminescence readout.I na ddition to the plasmodial and human A3pA hydrolases, we included PFE1035c (PfAp4Aase), which converts Ap4A into ATPa nd AMP.P fAp4Aase has recently been proposed as ap otential antimalaria targeta nd was included in this study as ac ontrol. [8] Indeed, we observed A3pA hydrolase activity for recombinant PfAp3Aase in vitro:t he Michaelis-Menten constant (K m )w as determined for the enzyme PfAp3Aase with the preferred substrate Ap3A. The observed K m value of 10.2 mm Ap3A is similar to the values described for the yeast (5.3 mm)a nd the human (1.2 mm)h omologues.…”

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confidence: 70%

Gift from Nature: Cyclomarin A Kills Mycobacteria and Malaria Parasites by Distinct Modes of Action

et al. 2015

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Malaria continues to be one of the most devastating human diseases despite many efforts to limit its spread by prevention of infection or by pharmaceutical treatment of patients. We have conducted a screen for antiplasmodial compounds by using a natural product library. Here we report on cyclomarin A as a potent growth inhibitor of Plasmodium falciparum and the identification of its molecular target, diadenosine triphosphate hydrolase (PfAp3Aase), by chemical proteomics. Using a biochemical assay, we could show that cyclomarin A is a specific inhibitor of the plasmodial enzyme but not of the closest human homologue hFHIT. Co-crystallisation experiments demonstrate a unique binding mode of the inhibitor. One molecule of cyclomarin A binds a dimeric PfAp3Aase and prevents the formation of the enzyme⋅substrate complex. These results validate PfAp3Aase as a new drug target for the treatment of malaria. We have previously elucidated the structurally unrelated regulatory subunit ClpC1 of the ClpP protease as the molecular target of cyclomarin A in Mycobacterium tuberculosis. Thus, cyclomarin A is a rare example of a natural product with two distinct and specific modes of action.

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“…Hence, signaling mediated by these molecules within red blood cells is of special interest in malaria. [7] Purinergic signaling has been shown to play role in parasite invasion. [8] The malaria parasite enzyme Ap4A hydrolase regulates levels of signaling molecules like Ap4A by hydrolyzing them to adenosine triphosphate and adenosine monophosphate.…”

Section: Introductionmentioning

confidence: 99%

Untitled

2017

AJP

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Aims: Malaria remains a worldwide health problem. Resistance to antimalarial drugs by the Plasmodium falciparum malaria parasite is posing a serious impediment in malaria control program. Hence continuous efforts on the development of new antimalarial are required. Being an important enzyme in parasite physiology diadenosine tetraphosphate (Ap4A) hydrolase may prove a good target for antimalarial drugs. The aim of this study was to explore the inhibitory potential of some sulfonamide derivatives against malaria parasite Ap4A hydrolase. Materials and Methods: The molecular three-dimensional structural data of malaria parasite Ap4A hydrolase were obtained from protein databank (PDB ID: 5CFI) and used as a drug target. The molecular docking approach was employed to find out the in silico inhibitory potential of the sulfonamide derivatives against Ap4A hydrolase and their relative stabilities were studied. Results and Discussion: All sulfonamide derivatives showed good binding affinity against the target protein. The binding free energy of compound 4-amino-N-(quinoxalin-2yl) benzenesulfonamide S6 (−8.43 Kcal/mole) showed it to be the most optimal sulfonamide derivative as inhibitor for Ap4A hydrolase enzyme. Conclusions: Evaluation of binding affinities using free energy simulations allowed establishing that sulfonamides having quinoxaline moiety is the highest quality compound of the series. The findings attained through this study on the molecular interaction mode of sulfonamide derivatives, and Ap4A hydrolase enzyme can be considered for further in vitro and in vivo validation for designing new potential antimalarial drugs.

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Molecular Characterization and Mutational Analysis of Recombinant Diadenosine 5′,5″-P<sup>1</sup>,P<sup>4</sup>-Tetraphosphate Hydrolase from <i>Plasmodium falciparum</i>

Cited by 6 publications

References 18 publications

Structural and functional attributes of malaria parasite diadenosine tetraphosphate hydrolase

Structural and functional attributes of malaria parasite diadenosine tetraphosphate hydrolase

Gift from Nature: Cyclomarin A Kills Mycobacteria and Malaria Parasites by Distinct Modes of Action

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