Abstract
Dissemination of drug-resistant malaria parasites represents one of the most important public health problems; therefore, the development of new antimalarial compounds is required. Cyclic AMP-dependent protein kinase is implicated in numerous cellular processes and an essential role for this enzyme has also been reported in the intraerythrocytic growth of the malaria parasite. The cAMP-dependent protein kinase from Plasmodium falciparum (PfPKA) plays an important role in the parasite life cycle and represents an attractive target for the development of antimalarial drugs. In this work, a recombinant PfPKA catalytic subunit (PfPKAc) was over-expressed in Escherichia coli and successfully purified using a two-step chromatographic process. The enzymatic properties of the recombinant PfPKAc were then determined using a sensitive fluorogenic assay suitable for biochemical characterization and inhibitor screening. This work provides new insights on the study of PfPKAc that will contribute to future investigations of the parasite cAMP signaling pathway and to high-throughput screening of specific malarial PKA inhibitors.
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Acknowledgements
The authors would like to thank PhD Taudon Nicolas, Php Pradines Bruno and Miss Dumoulin Chloé for critical reading of the manuscript and Dr. Desplans Jérôme for scientific discussion. This work was funded by the French Armed Forces Medical Service and the French “Délégation Générale pour l’Armement” (contrat d’objectif 06co008). The opinions and assertions contained herein are those of the authors and are not to be construed as official or reflecting the views of the French Armed Forces Medical Service or the French Army at large. All authors declare that they have no conflict of interest
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Wurtz, N., Pastorino, B., Almeras, L. et al. Expression and biochemical characterization of the Plasmodium falciparum protein kinase A catalytic subunit. Parasitol Res 104, 1299–1305 (2009). https://doi.org/10.1007/s00436-008-1327-3
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DOI: https://doi.org/10.1007/s00436-008-1327-3