State and activity of protein kinase C in postischemic reperfused liver.
We have studied the activity and the phorbol-binding capacity of protein kinase C (PKC) in subcellular fractions, as well as the relative amount of the enzyme protein in rat livers reperfused after severe nonnecrogenic ischemia. Ischemia causes a significant decrease in PKC phosphotransferase activity in both membranes and cytosol which lasts long after the reestablishment of the blood flow. The phorbol-binding capacity of the membrane fraction shows the same behavior. The amount of PKC protein decreases during ischemia (-25%) but returns to normal after reperfusion more promptly than activity and binding capacity, suggesting that PKC resynthesized in postischemic livers is either functionally defective or incapacitated by unsuitable conditions of the environment. We have also measured the contents of some lipids that may influence PKC activity in the cell. During ischemia and reperfusion there is a significant increase in the content of 1,2-diacylglycerol (DAG), which is the physiological activator of PKC, but under the conditions occurring in the ischemic/postischemic livers DAG apparently cannot bind to the enzyme and fulfill its function. Total phospholipids, phosphatidylcholine, and phosphatidylethanolamine, which significantly decrease at 60 min of ischemia, return to normal levels 1 hr after reperfusion.
Piccoletti
R
R
Istituto di Patologia Generale, Università degli Studi di Milano, Italy.
Bendinelli
P
P
Arienti
D
D
Bernelli-Zazzera
A
A
D000818:Animals; D002458:Cell Fractionation; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004075:Diglycerides / Q000032:analysis; D015151:Immunoblotting; D008055:Lipids / Q000032:analysis; D008099:Liver / Q000098:blood supply*; D008297:Male; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D010704:Phorbols / Q000378:metabolism; D010713:Phosphatidylcholines / Q000032:analysis; D010714:Phosphatidylethanolamines / Q000032:analysis; D010743:Phospholipids / Q000032:analysis; D011493:Protein Kinase C / Q000032:analysis / Q000378:metabolism* / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D015427:Reperfusion Injury / Q000201:enzymology*; D014316:Tritium
C011439:1,2-diacylglycerol; D004075:Diglycerides; D008055:Lipids; D010704:Phorbols; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D014316:Tritium; D015240:Phorbol 12,13-Dibutyrate; D011493:Protein Kinase C