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Background  cAMP-dependent protein kinase (PKA) has been implicated in the asexual stage of the  Toxoplasma gondii  life cycle through assaying the effect of a PKA-specific inhibitor on its growth rate. Since inhibition of the host cell PKA cannot be ruled out, a more precise evaluation of the role of PKA, as well as characterization of the kinase itself, is necessary.    Methodology/Principal Finding  The inhibitory effects of two PKA inhibitors, H89, an ATP-competitive chemical inhibitor, and PKI, a substrate-competitive mammalian natural peptide inhibitor, were estimated. In the  in vitro  kinase assay, the inhibitory effect of PKI on a recombinant  T. gondii  PKA catalytic subunit (TgPKA-C) was weaker compared to that on mammalian PKA-C. In a tachyzoite growth assay, PKI had little effect on the growth of tachyzoites, whereas H89 strongly inhibited it. Moreover,  T. gondii  PKA regulatory subunit (TgPKA-R)-overexpressing tachyzoites showed a significant growth defect.    Conclusions/Significance  Our data suggest that PKA plays an important role in the growth of tachyzoites, and the inhibitory effect of substrate-competitive inhibitor PKI on  T. gondii  PKA was low compared to that of the ATP competitive inhibitor H89.

Identification of Toxoplasma gondii cAMP Dependent Protein Kinase and Its Role in the Tachyzoite Growth