title: Expression and biochemical characterization of the Plasmodium falciparum protein kinase A catalytic subunit.
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.
authors.affiliation: UMR MD3 (UR3P), Relation hôte-parasites-Pharmacologie et Thérapeutique, Institut de Médecine Tropicale du Service de Santé des Armées, Marseille, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Wurtz
authors.forename: Boris
authors.initials: B
authors.lastname: Pastorino
authors.forename: Lionel
authors.initials: L
authors.lastname: Almeras
authors.forename: Sébastien
authors.initials: S
authors.lastname: Briolant
authors.forename: Claude
authors.initials: C
authors.lastname: Villard
authors.forename: Daniel
authors.initials: D
authors.lastname: Parzy
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002853:Chromatography, Liquid / Q000379:methods; D003001:Cloning, Molecular; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
chemical_list: D017868:Cyclic AMP-Dependent Protein Kinases