Expression and biochemical characterization of the Plasmodium falciparum protein kinase A catalytic subunit.
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.
Wurtz
Nathalie
N
UMR MD3 (UR3P), Relation hôte-parasites-Pharmacologie et Thérapeutique, Institut de Médecine Tropicale du Service de Santé des Armées, Marseille, France.
Pastorino
Boris
B
Almeras
Lionel
L
Briolant
Sébastien
S
Villard
Claude
C
Parzy
Daniel
D
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
D017868:Cyclic AMP-Dependent Protein Kinases