Heterogeneity of rat liver cytosol casein kinase 2. Association between the alpha/alpha' -subunits of casein kinase 2 and the phosphorylatable protein pp49.
Casein kinase 2 activity could be resolved into three peaks by chromatography on DEAE-Sepharose. The peak eluted at high salt concentrations (casein kinase 2b) showed molecular and kinetic properties typical of the heterotetramer composed of alpha-(or alpha'-) and beta-subunits. In contrast, the peak that was eluted at low salt concentrations (casein kinase 2a) contained no beta-subunit but a phosphorylatable protein of 49 kDa (pp49), in addition to the alpha/alpha'-subunits. The presence of alpha/alpha'/alpha"-subunits in preparations of casein kinases 2a and 2b was confirmed by immunological assays. Casein kinase 2a had low specific activity and a very high apparent Km for beta-casein. The peak eluted at intermediate ionic strength contained the alpha/alpha'-subunits and variable amounts of beta-subunit and pp49, and had kinetic properties intermediate between those of casein kinases 2a and 2b. Experiments based on heat inactivation, inhibition by low concentrations of heparin and ability to use GTP as substrate suggested that phosphorylation of pp49 was catalysed by the alpha/alpha'-subunits of casein kinase 2. No similarities were observed in the phosphopeptide maps of pp49 and beta-subunit. These results show that the alpha/alpha'-subunits of rat liver cytosol casein kinase 2 can form complexes not only with the beta-subunit but also with pp49, and that the complexes containing pp49 have a reduced affinity for the exogenous protein substrate beta-casein.
Molina
E
E
Departament de Bioquímica i Biologia Molecular-Unitat de Bioquímica Facultat de Ciències, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Plana
M
M
Itarte
E
E
D000818:Animals; D015153:Blotting, Western; D047388:Casein Kinases; D002364:Caseins / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D006493:Heparin / Q000494:pharmacology; D006358:Hot Temperature; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D008970:Molecular Weight; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats
D002364:Caseins; D046911:Macromolecular Substances; D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D006493:Heparin; D011494:Protein Kinases; D047388:Casein Kinases