Further characterization of galloyl pedunculagin as an effective autophosphorylation inhibitor of C-kinase in vitro.

The inhibitory effect of galloyl pedunculagin (GP) isolated from Platycarya strobilacea on the activity and autophosphorylation of Ca(2+)- and phospholipid-dependent protein kinase (C-kinase) was examined in vitro. It was found that (i). GP inhibited the activity (phosphorylation of complement C3 from guinea pig) of C-kinase alpha (rat brain) in a dose-dependent manner with an ID(50) of approx. 0.12 micro M (ii). GP at lower doses (ID(50)=approx. 6 nM) inhibited autophosphorylation of C-kinase alpha; and (iii). the GP-induced inhibition of autophosphorylation of C-kinase alpha and its enzyme activity was a manner non-competitive to ATP. Similar inhibitory effect of GP on autophosphorylation of recombinant human C-kinase eta (rhC-kinase eta) and its phosphorylating activity was observed. These results suggest that GP is an effective autophosphorylation inhibitor of these two C-kinase isoforms (alpha and eta) in vitro. In addition, the CD analysis suggests that the proline-containing six amino acid residues (PVLTPP) including a threonine residue (autophosphorylation site) at the C-terminal region (positions 635-640) of C-kinase alpha may be one of the GP-binding sites.