Skip to main content
SpringerLink
Log in

Inactivation of protein kinase C in rat liver during late hypoglycemic phase of sepsis

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Changes in protein kinase C (PKC) (calcium- and phospholipid-dependent protein kinase) activity in rat liver during different metabolic phases of sepsis were studied. Sepsis was induced by cecal ligation and puncture (CLP). Experiments were divided into three groups: control, early sepsis, and late sepsis. Early and late sepsis refers to those animals sacrificed at 9 and 18 h, respectively, after CLP. Hepatic PKC was extracted and partially purified by ammonium sulfate fractionation and DEAE-cellulose chromatography. PKC activity was assayed based on the rate of incorporation of 32p from [γ-32P]ATP into histone. The results show that during early sepsis, both membrane-associated and cytosolic PKC activities remained relatively unaltered. During late sepsis, membrane-associated PKC was unaffected while cytosolic PKC activity was decreased by 19.5-34.4%. Kinetic analysis of the data on cytosolic PKC during late phase of sepsis reveals that the Vmax values for ATP, histone, Ca2+, phosphatidylserine, and diacylglycerol were decreased by 23.4, 22.1, 19.5, 25, and 34.4%, respectively, with no changes in their Km values. These data indicate that cytosolic PKC activity was inactivated in rat liver during late hypoglycemic phase of sepsis. Since PKC-mediated phosphorylation plays an important role in regulating hepatic glucose metabolism, an inactivation of cytosolic PKC may contribute to the development of hypoglycemia during late phase of sepsis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Blobe GC, Stribling S, Obeid LM, Hannun YA: Protein kinase C isoenzymes: Regulation and function. Cancer Surv 27: 213–248, 1996

    Google Scholar 

  2. Patel TB: Stimulation of hepatic glycogenolysis by phorbol 12-myristate 13-acetate. Biochem J 241: 549–554, 1987

    Google Scholar 

  3. Kimura S, Nagasaki K, Adachi I, Yamaguchi K, Fujiki H, Abe K: Stimulation of hepatic glycogenolysis by 12-0-tetradecanoylphorbol-13-acetate (TPA) via a calcium requiring process. Biochem Biophys Res Commun 122: 1057–1064, 1984

    Google Scholar 

  4. Inaba H, Araki M, Numai T: Modulation of protein kinase C alters hemodynamics and metabolism in the isolated liver in fed and fasted rats. J Hepatol 19: 475–484, 1993

    Google Scholar 

  5. Gali RR, Pugazhenthi S, Khandelwal RL: Reciprocal effects of the protein kinase C inhibitors staurosporine and H-7 on the regulation of glycogen synthase and phosphorylase in the primary culture of hepatocytes. Metabolism 42: 1475–1480, 1993

    Google Scholar 

  6. Roach PJ, Goldman M: Modification of glycogen synthase activity in isolated rat hepatocytes by tumor-promoting phorbol esters: Evidence for differential regulation of glycogen synthase and phosphorylase. Proc Natl Acad Sci USA 80: 7170–7172, 1983

    Google Scholar 

  7. Blackmore PF, Strickland WG, Bocckino SB, Exton JH: Mechanism of hepatic glycogen synthase inactivation induced by Ca2+-mobilizing hormones. Studies using phospholipase C and phorbol myristate acetate. Biochem J 237: 235–242, 1986

    Google Scholar 

  8. Arino J, Guinovart JJ: Phosphorylation and inactivation of rat hepatocyte glycogen synthase by phorbol esters and mezerein. Biochem Biophys Res Commun 134: 113–119, 1986

    Google Scholar 

  9. Fain JN, Li SY, Litosch I, Wallace M: Synergistic activation of rat hepatocyte glycogen phosphorylase by A23187 and phorbol ester. Biochem Biophys Res Commun 119: 88–94, 1984

    Google Scholar 

  10. McGilvery RW: Storage of glucose as glycogen. In: RW McGilvery (ed). Biochemistry, A Functional Approach. W.B. Saunders Company, Philadelphia, 1979, pp 483–510

    Google Scholar 

  11. Liu MS, Sharma C: Glycolytic enzyme activities in normal and diabetic dog livers during endotoxic shock. Am J Physiol 240: R10–R15, 1981

    Google Scholar 

  12. Hwang TL, Lau YT, Chen MP, Tang C, Liu MS: Biphasic intracellular redistribution of α1-adrenergic receptors in rat liver during sepsis. Am J Physiol 265: R385–R391, 1993

    Google Scholar 

  13. Liu MS, Kang GF: Liver glycogen metabolism in endotoxin shock. I. Endotoxin administration decreases glycogen synthase activities in dog livers. Biochem Med Metabol Biol 37: 61–72, 1987

    Google Scholar 

  14. Liu MS, Kang GF: Liver glycogen metabolism in endotoxin shock. II. Endotoxin administration increases glycogen phosphorylase activities in dog liver. Biochem Med Metabol Biol 37: 73–80, 1987

    Google Scholar 

  15. Inaba H, Filkins JP: Augmentation of endotoxin lethality and glucose dyshomeostasis by phorbol ester. Am J Physiol 260: R494–R502, 1991

    Google Scholar 

  16. Inaba, H, Pilkins JP: Antagonism of endotoxic glucose dyshomestasis by protein kinase C inhibitors. Am J Physiol 261: R26–R31, 1991

    Google Scholar 

  17. Wichterman KA, Baue AK, Chaudry IH: Sepsis and septic shock – A review of laboratory models and a proposal. J Surg Res 29: 189–210, 1980

    Google Scholar 

  18. Wu LL, Tang C, Liu MS: Hyper and hypocardiodynamic states are associated with externalization and internalization, respectively, of α-adrenergic receptors in rat heart during sepsis. Shock 5: 318–323, 1997

    Google Scholar 

  19. Wise BC, Raynor RL, Kuo JF: Phospholipid-sensitive Ca2+-dependent protein kinase from heart. I. Purification and general properties. J Biol Chem 257: 8481–8488, 1982

    Google Scholar 

  20. Fabiato A: Computer programs for calculating total from specified free or free from specified total ionic concentrations in aqueous solutions containing multiple metals and ligands. Meth Enzymol 157: 378–417, 1988

    Google Scholar 

  21. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  22. Chan KM, Junger KD: Calcium transport and phosphorylated intermediate of (Ca2+-Mg2+)-ATPase in plasma membranes of rat liver. J Biol Chem 258:4404–4410, 1983

    Google Scholar 

  23. Pavoine C, Lotersztajn S, Mallat A, Pecker F: The high affinity (Ca2+-Mg2+)-ATPase in liver plasma membrane is a Ca2+ pump: Reconstitution of the purified enzyme into phospholipid vesicles. J Biol Chem 262: 5113–5117, 1987

    Google Scholar 

  24. Evers C, Hugentobler G, Lester R, Gmaj P, Meier P, Murer H: ATP-dependent Ca2+ uptake and Ca2+-dependent protein phosphorylation in basolateral liver plasma membrane. Biochim Biophys Acta 939: 542–550, 1988

    Google Scholar 

  25. Monteith GR, Roufogalis BD: The plasma membrane calcium pump – a physiological perspective on its regulation. Cell Calcium 18: 459–470, 1995

    Google Scholar 

  26. Wright LC, Chen S, Roufogalis BD: Regulation of the activity and phosphorylation of the plasma membrane Ca2+-ATPase by protein kinase C in intact human erythrocytes. Arch Biochem Biophys 306: 277–284, 1993

    Google Scholar 

  27. Furukawa KI, Tawada Y, Shigekawa M: Protein kinase C activation stimulates plasma membrane Ca2+ pump in cultured vascular smooth muscle cells. J Biol Chem 264: 4844–4849, 1989

    Google Scholar 

  28. Lagast H, Pozzan T, Waldvogel FA, Lew PD: Phorbol myristate acetate stimulates ATP dependent calcium transport by the plasma membrane of neutrophils. J Clin Invest 73: 878–883, 1984

    Google Scholar 

  29. Lau YT, Hwang TL, Chen MF, Liu MS: Calcium transport by rat liver plasma membranes during sepsis. Circ Shock 38: 238–244, 1992

    Google Scholar 

  30. Kang YH, Mekenna T, Watson LP, Williams R, Holt M: Cytochemical changes in hepatocytes of rats with endotoxemia or sepsis: Localization of fibronectin, calcium and enzymes. J Histochem Cytochem 36: 665–678, 1988

    Google Scholar 

  31. Sayeed MM: Alterations in cellular Ca2+ regulation in the liver in endotoxic shock. Am J Physiol 250: R884–R891, 1986

    Google Scholar 

  32. Newton AC: Protein kinase C: Structure, function, and regulation. J Biol Chem 270: 28495–28498, 1995

    Google Scholar 

  33. Liu JP: Protein kinase C and its substrate. Mol Cell Endocrinol 116: 1–29, 1996

    Google Scholar 

  34. Nishizuka Y: Intracellular signalling by hydrolysis of phospholipids and activation of protein kinase C. Science 258: 607–614, 1992

    Google Scholar 

  35. Go M, Sekiguchi K, Nomura H, Kikkawa U, Nishizuka Y: Further studies on the specificity of diacylglycerol for protein kinase C activation. Biochem Biophys Res Commun 144: 598–605, 1987

    Google Scholar 

  36. Ford DA, Miyake R, Glaser PE, Gross RW: Activation of protein kinase C by naturally occurring ether-linked diglycerides. J Biol Chem 264: 13818–13824, 1989

    Google Scholar 

  37. Nishizuka Y: Protein kinase C and lipid signalling for sustained cellular responses. FASEB J 9: 484–496, 1995

    Google Scholar 

  38. Nishizuka Y: The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature 334: 661–665, 1988

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Physiology, Graduate Institute of Medicine, Kaohsiung Medical College, Kaohsiung, Taiwan

    Chin Hsu, Hsiao-Ching Jao, Shaw-Lang Yang & Hseng-Kuang Hsu

  2. Department of Pharmacological and Physiological Science, Saint Louis University School of Medicine, St. Louis, Missouri, 63104, USA

    Maw-Shung Liu

Authors
  1. Chin Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hsiao-Ching Jao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shaw-Lang Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hseng-Kuang Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Maw-Shung Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hsu, C., Jao, HC., Yang, SL. et al. Inactivation of protein kinase C in rat liver during late hypoglycemic phase of sepsis. Mol Cell Biochem 181, 181–189 (1998). https://doi.org/10.1023/A:1006853106320

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006853106320

Search

Navigation