zeta-Related protein kinase C in nuclei of nerve cells.
To determine whether or not PKC is present in the nuclei of nerve tissue we made use of biochemical and immunocytochemical techniques. A 219-fold purification of rabbit brain nuclear protein kinase C was achieved by sequential steps of Triton X-100 extraction of isolated nuclei, DEAE-cellulose, Butyl-toyopearl and hydroxylapatite chromatography. The major peak of protein kinase C activity was eluted from the hydroxylapatite column at the KPO4 concentration of 0.3 M. Both Ca2+ and Ptd Ser were required for stimulation of the enzyme. Immunoblot analysis revealed that the kinase fraction was immunoreactive with a polyclonal antibody, PC-zeta, that had been raised against a peptide synthesized according to the deduced sequence of rat zeta protein kinase C. Light-microscopy revealed strong immunoreactivity in the nuclei of Purkinje cells in cerebellum and pyramidal cells in the rat cerebral cortex. These observations suggest that a zeta-related protein kinase C is present in the nuclei of nerve cells.
Hagiwara
M
M
Department of Pharmacology, Nagoya University School of Medicine, Aichi, Japan.
Uchida
C
C
Usuda
N
N
Nagata
T
T
Hidaka
H
H
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D002118:Calcium / Q000502:physiology; D002467:Cell Nucleus / Q000201:enzymology*; D007120:Immunochemistry; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D009474:Neurons / Q000201:enzymology*; D011493:Protein Kinase C / Q000302:isolation & purification / Q000378:metabolism*; D011817:Rabbits; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D007527:Isoenzymes; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium