Partial purification and characterization of a calcium-dependent protein kinase in rice leaves.

A protein from rice leaves, which was partially purified by sequential chromatography on DE52, MONO-Q and Superose 12, presented calcium-dependent protein kinase (CDPK) activity. This protein kinase phosphorylated the substrate, histone III-S, in a Ca(2+)-dependent manner and the half-maximum concentration of Ca2+ to protein kinase activity (EC50) was 1 microM. This phosphorylation was independent of phosphatidylserine and a phorbol ester. The apparent M(r) of the protein kinase, as determined by phosphorylation in SDS-polyacrylamide gel containing histone III-S, was 45 k. This kinase was found to react differently from other protein kinases, such as protein kinase C from rat brain or CDPK from soybean leaves, owing to the absence of a phospholipid or phorbol ester dependency. CDPK phosphorylated three endogenous proteins as detected by in vitro phosphorylation on two-dimensional PAGE.