Protein kinase C located in rat liver nuclei. Partial purification and biochemical and immunochemical characterization.
In the rat liver homogenate, maximal protein kinase C activity was found at two calcium concentrations (1.75 and 3.5 mM). Subcellular fractionation of the liver homogenate revealed that the protein kinase C activity requiring 1.75 mM calcium was present only in the cytosolic and particulate subcellular fractions. The protein kinase C activity requiring 3.5 mM calcium concentration was mainly located in the rat liver nuclei preparation. About 19% of the liver homogenate protein kinase C activity requiring 3.5 mM calcium was present in the nuclei. Goat anti-rat brain protein kinase C antibodies revealed a single immunoreactive band at 80-82 kDa in the rat liver nuclear, particulate, or cytosolic fractions. Based on the ratio of plasma membrane marker enzyme activity determined in the nuclear preparation, the purity of the isolated nuclei was ascertained. Rat liver nuclear protein kinase C activity has been partially purified. The purification steps sequentially employed were Triton X-100 extraction of isolated nuclei, DEAE-cellulose chromatography, Phenyl-Superose, and Mono Q (fast protein liquid) chromatography. The final purification step revealed, by silver nitrate staining on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two protein bands at 80 and 66 kDa, respectively. These findings provide definitive data regarding the nuclear location of protein kinase C. The nuclear location of protein kinase C may lead to an understanding of the molecular pathway involved in signal transduction from the plasma membrane to the nucleus.
Masmoudi
A
A
Centre de Neurochimie, l'Institut National de la Sante, Strasbourg, France.
Labourdette
G
G
Mersel
M
M
Huang
F L
FL
Huang
K P
KP
Vincendon
G
G
Malviya
A N
AN
D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex / Q000032:analysis; D002118:Calcium / Q000494:pharmacology; D002458:Cell Fractionation / Q000379:methods; D002467:Cell Nucleus / Q000201:enzymology*; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D015151:Immunoblotting; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D011493:Protein Kinase C / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000201:enzymology
D000906:Antibodies; D000936:Antigen-Antibody Complex; D011493:Protein Kinase C; D002118:Calcium