Purification and characterization of casein kinase II from lactating rabbit mammary gland

1. Highly purified 200 kDa casein kinase II from rabbit lactating mammary gland (MG-CK II) was obtained by means of a new purification procedure consisting of one phosphocellulose and three Monó Q steps. 2. Its Km for ATP was 2.22 microM and 0.57 mg/ml and 0.13 mg/ml for partially dephosphorylated casein and phosvitin respectively. Stathmine was also suitable as substrate. 2-aminopurine and 6-dimethylaminopurine inhibited efficiently MG-CK II (Ki = 5 and 1 mM respectively). 3. MG-CK II autophosphorylated on its alpha-, alpha'- and beta-subunits. The beta-subunit autophosphorylation was enhanced in presence of exogenous substrate. Its modulation was highly dependent on ATP concentration. 4. The effects of basic compounds which affected dramatically the phosphorylation of dephosphorylated casein in presence of various ATP concentrations were reported.