Enhancement of protein kinase activity in the cytosol of regenerating rat liver: regulatory role of endogenous regucalcin.
The effect of regucalcin, a Ca2+-binding protein, on protein kinase activity in the cytosol of regenerating rat liver was investigated. Protein phosphorylation was significantly increased in the liver cytosol obtained at 6, 24, and 48 h after a partial hepatectomy (about 65%) in comparison with that of sham-operated rats. This increase was significantly inhibited by the addition of trifluoperazine (2x10(-5) M), staurosporine (10(-7) M) or genistein (10(-5) M), which is an inhibitor of protein kinases, in the reaction mixture. The presence of regucalcin (0.1-0.5 microM) caused a significant decrease in protein phosphorylation in the cytosol from normal and regenerating rat livers. The effect of regucalcin (0.5 microM) was completely abolished by the addition of anti-regucalcin monoclonal antibody (50 ng/ml). The elevation of protein phosphorylation in regenerating rat liver was significantly enhanced by the presence of anti-regucalcin monoclonal antibody (50 ng/ml). The effect of regucalcin in decreasing protein phosphorylation in the cytosol of regenerating rat liver was not seen in the presence of the antibody. The present study demonstrates that protein kinase activity, is enhanced in the cytosol of regenerating rat liver, and that endogenous regucalcin has an inhibitory role in the enhancement of protein phosphorylation by various protein kinases.
Yamaguchi
M
M
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, Shizuoka City 422-8526, Japan.
Katsumata
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D020013:Calcium Signaling; D002135:Calcium-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000494:pharmacology; D002265:Carboxylic Ester Hydrolases; D003600:Cytosol / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008115:Liver Regeneration / Q000187:drug effects / Q000502:physiology*; D008297:Male; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015238:Sulfotransferases
D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D015238:Sulfotransferases; C028152:alcohol sulfotransferase; D002265:Carboxylic Ester Hydrolases; C513325:Rgn protein, rat; D002118:Calcium