A protein kinase C inhibitory activity is present in rat brain homogenate

• Published in: FEBS letters Vol. 177; no. 1; pp. 36 - 40
• Main Authors: Schwantke, Nicole, Le Peuch, Christian J.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 05.11.1984; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; brain; Brain Chemistry; Calcium; Carrier Proteins - analysis; Chromatography, DEAE-Cellulose; Chromatography, Gel; Enzyme Inhibitors - analysis; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Histones - metabolism; Intracellular Signaling Peptides and Proteins; Kinase; Kinase inhibitor; Male; Phosphoinositide; Phospholipid; phospholipids; Phosphorylation; protein kinase; Protein Kinase C; protein kinase inhibitor; Protein Kinase Inhibitors; Rats; Rats, Inbred Strains; Transferases; Trypsin - metabolism; Phospholipid; Phosphoinositide; Calcium; Kinase; Kinase inhibitor; Vertebrata; Mammalia; Purification; Rat; Enzyme; Protein kinase; Rodentia; Brain (Vertebrata); Inhibitor
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(84)80976-X

The partial purification and characterization of (a) factor(s) from rat brain which inhibit(s) the activity of calcium and phospholipid-dependent protein kinase from the same tissue is described. This factor, present in 100000 × g rat brain homogenate supernatant, is inactivated upon treatment by trypsin and pepsin and is therefore assumed to be a protein. It was partially purified by ion-exchange chromatography on DEAEcellulose, ammonium sulfate precipitation and gel filtration. This inhibitor is not stable to heating at 70°C for 10 min, however partial renaturation of the inhibitory activity can be observed after incubation of the denatured inhibitor for 24 h at 4°C. It is precipitable by 10% trichloroacetic acid and by 2 M ammonium sulfate. It exhibits a Stokes radius of 20 Å by gel exclusion chromatography, corresponding to a molecular mass of 20 kDa assuming a globular shape. Kinetic analysis of the inhibition of calcium-phospholipid-dependent histone kinase activity indicates that the inhibitor is competitive with respect to the protein substrate. No change was observed in the kinetic values of the kinase for ATP, Ca 2+ and phospholipids.