Protein kinase activity associated with immunopurified p53 protein.
Enhanced protein phosphorylation seems to be characteristic for cell transformation. Viral or cellular oncogene products which are functionally implicated in cell transformation sometimes activate protein kinases, or they are protein kinases themselves. In the present paper we have shown that a protein kinase activity is tightly associated with immunopurified oncoprotein p53, either uncomplexed or in complex with SV40 large T antigen. Furthermore, we could demonstrate that the protein kinase associated with immunopurified p53 was independent of SV40 large T antigen. p53 in the immunocomplexes served as a substrate for this protein kinase. Phosphoamino acid analysis of in vitro phosphorylated p53 revealed a phosphorylation predominantly on serine residues similar to p53 phosphorylated in vivo. The use of different monoclonal antibodies did not reveal a total inhibition of the protein kinase activity. However, p53 precipitated with monoclonal antibodies which recognize a C-terminal domain, was phosphorylated in vitro to a lesser extent than p53 which was precipitated with monoclonal antibodies that recognize an N-terminal epitope. All subclasses of immunopurified p53 separable by sucrose density gradients or by sequential immunoprecipitation exhibited a protein kinase activity and served as substrates for this protein kinase. Moreover, a protein kinase activity was found to be associated with baculovirus expressed p53 which allows us to attribute this enzymatic activity more directly to p53.
Kraiss
S
S
Department of Biochemistry, University of Ulm, Federal Republic of Germany.
Barnekow
A
A
Montenarh
M
M
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015513:Oncogene Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D010750:Phosphoproteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D016159:Tumor Suppressor Protein p53
D000596:Amino Acids; D000945:Antigens, Differentiation, T-Lymphocyte; D015513:Oncogene Proteins; D010750:Phosphoproteins; D016159:Tumor Suppressor Protein p53; D011494:Protein Kinases