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A protein kinase C inhibitory activity is present in rat brain homogenate
Author(s) -
Schwantke Nicole,
Le Peuch Christian J.
Publication year - 1984
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(84)80976-x
Subject(s) - ammonium sulfate precipitation , chemistry , size exclusion chromatography , biochemistry , chromatography , trypsin , phospholipid , stokes radius , protein kinase a , enzyme , membrane
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 1 × 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.