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Calcium-dependent protein kinases (CDPKs) are important in calcium influx, triggering several biological processes in  Cryptosporidium  spp. As they are not present in mammals, CDPKs are considered promising drug targets. Recent studies have characterized  Cp CDPK1,  Cp CDPK3,  Cp CDPK4,  Cp CDPK5,  Cp CDPK6, and  Cp CDPK9, but the role of  Cp CPK2A remains unclear. In this work, we expressed recombinant  Cp CDPK2A encoded by the cgd2_1060 gene in  Escherichia coli  and characterized the biologic functions of  Cp CDPK2A using qRT-PCR, immunofluorescence microscopy, immuno-electron microscopy, and  in vitro  neutralization. The results revealed that  Cp CDPK2A protein was highly expressed in the apical region of sporozoites and merozoites and in macrogamonts. Monoclonal or polyclonal antibodies against  Cp CDPK2A failed to block the invasion of host cells. Among the 44 candidate inhibitors from molecular docking of  Cp CDPK2A, one inhibitor was identified as having a potential effect on both  Cryptosporidium parvum  growth and  Cp CDPK2A enzyme activities. These data suggest that  Cp CDPK2A may play some roles during the development of  C. parvum  and might be a potential drug target against cryptosporidiosis.

Characterization of Calcium-Dependent Protein Kinase 2A, a Potential Drug Target Against Cryptosporidiosis