Developmental changes of protein substrates of Ca2+/calmodulin-dependent protein kinase II in the rat forebrain

We previously reported that the level of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) alpha and beta proteins increases with postnatal age. In the present study, we investigated the developmental changes in whole protein substrates of CaM kinase II as compared with those of cAMP-dependent protein kinase (A-kinase) in the rat forebrain. Protein substrates were phosphorylated with [gamma-33P]ATP, and analysed by two-dimensional gel electrophoresis. More than 50 substrates for CaM kinase II were found in the soluble and particulate fractions. The phosphorylation level of more than 15 substrates increased in the particulate fraction during development. Similarly, that of more than 3 substrates increased in the soluble fraction. Some substrates for A-kinase also increased during development, although some decreased. These findings suggest that the expression of some substrates is regulated during development and that the phosphorylation reaction involves the regulation of neuronal development.