Adenosine 3′,5′-monophosphate-dependent protein kinase(s) in diploid and SV40 transformed human fibroblasts
| Autor: | Thomas B. Miller, Reginald W. Butcher, Keith P. Ray |
|---|---|
| Rok vydání: | 1979 |
| Předmět: |
Biophysics
Protamine Kinase Simian virus 40 Mitogen-activated protein kinase kinase Biochemistry Cell Line MAP2K7 Glycogen phosphorylase Cyclic AMP Humans Protein kinase A Molecular Biology MAPK14 biology Kinase Prostaglandins E Cyclin-dependent kinase 2 Cyclin-dependent kinase 3 Diploidy Molecular biology Enzyme Activation Kinetics Cell Transformation Neoplastic biology.protein Protein Kinases |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - General Subjects. 584:181-195 |
| ISSN: | 0304-4165 |
| DOI: | 10.1016/0304-4165(79)90263-0 |
| Popis: | Cyclic AMP-dependent protein kinases (EC 2.7.1.37; ATP:protein phosphotransferase) in the human diploid fibroblast WI-38 and an SV40-transformant WI-38-VA13-2RA (VA13) have been compared on the basis of their concentrations in cells, isoenzyme composition and susceptibility to hormonal activation. In high population density cultures, total soluble cyclic AMP-dependent kinase activities measured with histone were essentially the same in WI-38 and VA13. Two soluble protein kinase forms separated by chromatography on DEAE-cellulose were present in both cell lines. The concentration of cyclic AMP required for half-maximal activation of both enzyme forms was 10-30 nM. Overall kinase stimulation was greater for the Peak I enzymes. Kinase activation induced in the presence of 0.5 M KCl was more rapid and complete for the Peak I enzymes. Under conditions which elevated the concentration of cyclic AMP in WI-38 and VA13 cells the activities of the soluble histone kinases were increased. Incubation of the cells with either of 5.7 micronM prostaglandin E1 or 1 micronM isopropylnorepinephrine induced complete activation of the cyclic AMP-dependent histone kinases within 5 min and maintained the effect for 20 min. When intracellular cyclic AMP levels were raised by prostaglandin E1, activation of glycogen phosphorylase (assayed-AMP) suggested that this enzyme cascade involving cyclic AMP-dependent protein kinase(s) was intact and responsive in both cell lines. |
| Databáze: | OpenAIRE |
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