Lack of translocation of protein kinase C from the cytosol to the membranes in vasopressin-stimulated hepatocytes.
The ability of Ca2(+)-mobilizing hormones to promote changes in the subcellular distribution of protein kinase C (PKC) was studied in isolated hepatocytes. In recently isolated cells the distribution of PKC between the soluble and particulate fractions was 47 and 53% respectively. Exposure of the hepatocytes to 100 nM-vasopressin produced an increased phosphoinositide turnover, as reflected by the changes in the concentrations of inositol trisphosphate and Ca2+, and in glycogen phosphorylase a activity. However, the distribution of both PKC activity and [3H]phorbol dibutyrate binding between the cytosol and the membranes remained unchanged under these conditions. To determine the threshold values of the concentrations of Ca2+ and diacylglycerol required to produce a redistribution of PKC, the hepatocytes were treated with the Ca2+ ionophore ionomycin, and with permeant diacylglycerol derivatives. Hepatocytes incubated in the presence of 100 nM-vasopressin required concentrations of Ca2+ 2.5 times those produced physiologically by the hormone to produce translocation of PKC from the cytosol to the membranes. These studies suggest that, at least in hepatocytes, activation of PKC in response to Ca2(+)-mobilizing hormones involves only the pre-existent membrane-bound enzyme without affecting the soluble enzyme.
Díaz-Guerra
M J
MJ
Instituto de Bioquímica, Facultad de Farmacia, Universidad Complutense, Madrid, Spain.
Boscá
L
L
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology*; D004075:Diglycerides / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D008297:Male; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism; D010762:Phosphorylase a / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D014667:Vasopressins / Q000494:pharmacology*
D004075:Diglycerides; D010716:Phosphatidylinositols; D014667:Vasopressins; D015240:Phorbol 12,13-Dibutyrate; D015759:Ionomycin; C038237:1-oleoyl-2-acetylglycerol; D010762:Phosphorylase a; D011493:Protein Kinase C; D002118:Calcium