title: Protein phosphorylation in isolated hepatocytes of septic and endotoxemic rats.
abstract: The purpose of this study was to investigate possible alterations induced by sepsis and endotoxicosis in the late phase of Ca2+-dependent signaling in rat liver. Hepatocytes isolated from septic or chronically endotoxin (ET)-treated rats were labeled with [32P]H3PO4 and stimulated with various agents. Proteins were resolved by one-dimensional polyacrylamide gel electrophoresis and autoradiographed. Vasopressin (VP)- and phenylephrine (PE)-induced responses were attenuated in both septic and ET-treated rats for cytosolic and membrane proteins compared with their respective controls. Glucagon and 12-O-myristate phorbol-13-acetate (TPA) affected only the phosphorylation of membrane proteins. Glucagon-induced changes in the phosphorylation of membrane proteins were affected by both sepsis and endotoxicosis, whereas TPA-stimulated phosphorylation was lowered only in endotoxicosis. Response to the Ca2+ ionophore A23187 was depressed in septic rats for cytosolic proteins. The phosphorylation of two cytosolic proteins, i.e., 93 and 61 kDa (previously identified as glycogen phosphorylase and pyruvate kinase, respectively), in response to VP, PE, and A23187 was severely impaired by endotoxicosis and sepsis. TPA did not affect the phosphorylation state of these two proteins. The results show that sepsis and endotoxicosis produce perturbations of the phosphorylation step in Ca2+ transmembrane signaling. Such changes can explain alterations of glycogenolysis and gluconeogenesis associated with sepsis and endotoxicosis.
authors.affiliation: Department of Physiology, Louisiana State University Medical Center, New Orleans 70112.
authors.forename: I V
authors.initials: IV
authors.lastname: Deaciuc
authors.forename: J A
authors.initials: JA
authors.lastname: Spitzer
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002469:Cell Separation; D004072:Digitonin / Q000494:pharmacology; D004731:Endotoxins / Q000097:blood*; D007239:Infections / Q000378:metabolism* / Q000473:pathology; D008099:Liver / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D008297:Male; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013268:Stimulation, Chemical
chemical_list: D004731:Endotoxins; D010761:Phosphorus Radioisotopes; D011506:Proteins; D000255:Adenosine Triphosphate; D004072:Digitonin