The analysis of role of calcium-dependent protein kinase in the mechanism of parasitism of malarial parasites

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In 1994 : Polymerase chain reaction (PCR) was carried out against P.yoelii genomic DNA with oligonucleotide primers which were designed from highly conserved regions among calcium-dependent protein kinases (protein kinase C and Ca^<2+>/calmodulin-dependent protein kinase) of a wide range of organisms and a PCR product was yielded. P.yoelii genomic DNA library was screened using this PCR product as a primer. Sequencing of the insert of a positive clone revealed that it contained a gene which codes a protein kinase.In 1995 : The comparison of the primary structure of the parasite protein kinase with those of various kinds of protein kinases of other organisms showed that the parasite protein belongs to cAMP-dependent protein kinase catalytic subunit family members. In situ gel assay of P.yoelii proteins showed that there are at least 8 kinds of protein kinases of P.yoelii. And the calcium-dependent protein kinase showed the highest activity from late trophozoite stage to schizont stage. The protein kinase which is more activated in the presence of EGTA showed the highest activity from schizont stage to early trophozoite stage. The study of the effect of various inhibitors of protein kinases on the growth of P.falciparum in vitro demonstrated that inhibitors of calcium-dependent protein kinases (protein kinase C and Ca^<2+>/calmodulin-dependent protein kinase) inhibit the growth of P.falciparum.