Skip to main content
Log in

Atrial natriuretic factor inhibits autophosphorylation of protein kinase C and A 240-kDa protein in plasma membranes of bovine adrenal glomerulose cells: Involvement of cGMP-dependent and independent signal transduction mechanisms

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

Abstract

To clarify the intracellular signalling mechanisms of atrial natriuretic factor (ANF), we studied its effect on protein phosphorylation in plasma membranes of bovine adrenal cortical cells. ANF (1×10−7 M) inhibited phosphorylation of the 78-kDa protein kinase C (PKC) and a 240-kDa protein in specific manner. In parallel experiments, cGMP (0.5 mM) inhibited phosphorylation of only the 78-kDa PKC but it did not affect phosphorylation of the 240-kDa protein. Phosphorylation of the 78-kDa PKC was enhanced in a Ca2+-/phospholipid-dependent manner. However, after prolonged preincubation of plasma membranes with Ca2+ (0.5 mM), the incorporation of32P-radioactivity rapidly decreased in the 78-kDa PKC and subsequently increased in the 45- and 48-kDa protein bands due to Ca2+-dependent proteolytic degradation of 78-kDa PKC. Polyclonal antibodies against brain PCK were used to immunoblot and immunoprecipitate the 78-kDa PKC. Preincubation of plasma membranes with Ca2+ for varying times, followed by immunoblotting revealed a gradual loss of the immunoreactive 78-kDa PKC band in a time-dependent manner. Immunoprecipitation of phosphorylated 78-kDa PKC in plasma membranes showed that its phosphorylation was significantly inhibited in the presence of ANF as compared to control membranes, phosphorylated in the absence of ANF. The results in this present study document a new signal transduction mechanism of ANF at molecular level which possibly involves dephosphorylation of the 78-kDa PKC and a 240-kDa protein in a cGMP-dependent and-independent manner in bovine adrenal glomerulosa cell membranes. (Mol Cell Biochem141: 103–111, 1994)

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

Access this article

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. deBold AJ, Borenstein HB, Veress AT and Sonnenberg H: A rapid and potent natriuretic response to intravenous injection of atrial myocardial extracts in rats. Life Sci 28: 89–94, 1981

    Google Scholar 

  2. Inagami T: Atrial natriuretic factor. J Biol Chem 264: 3043–3046, 1989

    Google Scholar 

  3. Brenner BM, Ballerman BJ, Gunning ME and Zeidel ML: Diverse biological actions of atrial natriuretic peptide. Physiol Rev 70: 665–699, 1990

    Google Scholar 

  4. Rozenzweig A and Seidman CE: Atrial natriuretic factor and related peptide hormone. Ann Rev Biochem 60: 229–255, 1991

    Google Scholar 

  5. Maack T: Receptors for atrial natriuretic factor. Ann Rev Physiol 54: 11–27, 1992

    Google Scholar 

  6. Leitman DC, Andresen JW, Kuno T, Kamisaki Y, Chang J-K and Murad F: Identification of multiple binding sites for atrial ntriuretic factor by affinity crosslinking in cultured endothelial cells. J Biol Chem 261: 11650–11655, 1986

    Google Scholar 

  7. Scarborough RM, Schenk DB, McEnroe GA, Arfsten A, Kang L-L, Schwartz K and Lewicki JA: Truncated atrial natriuretic peptide analogs: Comparison between receptor binding and stimulation of cyclic GMP accumulation in cultured vascular smooth muscle cells. J Biol Chem 261: 12960–12964, 1986

    Google Scholar 

  8. Leitman DC, Andressen JW, Catalano RM, Waldman SA, Tuan JJ and Murad F: Atrial natriuretic peptide binding, cross-linking and stimulation of cGMP accumulation and particulate guanylate cyclase activity in cultured cells. J Biol Chem 263: 3720–3728, 1988

    Google Scholar 

  9. Pandey KN, Pavlou SN and Inagami T: Identification and characterization of three distinct atrial natriuretic factor receptors: Evidence for tissue specific heterogeneity of receptor subtypes in vascular smooth muscle, kidney tubular epithelium and Leydig tumor cells by ligand binding, photoaffinity labeling and tryptic proteolysis. J Biol Chem 263: 13406–13413, 1988

    Google Scholar 

  10. Garbers DL: Guanylate cyclase, a cell surface receptor. J Biol Chem 264: 9103–9106, 1989

    Google Scholar 

  11. Lowe DG, Chang M-S, Hellmiss R, Chen E, Singh S, Garbers DL and Goeddel DV: Human atrial natriuretic peptide receptor defines a new paradigm for second messenger signal transduction. EMBO J 8: 1377–1384, 1989

    Google Scholar 

  12. Chang M-S, Lowe DG, Lewis M, Hellmiss R, Chen E and Goeddel DV: Differential activation by atrial and brain natriuretic peptides of two different receptor guanylate cyclase. Nature 341: 68–72, 1989

    Google Scholar 

  13. Chinkers M, Garbers DL, Chang M-S, Lowe DG, Chin H, Goeddel DV and Schulz S: A membrane form of guanylate cyclase is an atrial natriuretic peptide receptor. Nature 338: 78–83, 1989

    Google Scholar 

  14. Schulz S, Singh S, Bellet RA, Singh S, Tubb DJ, Chen H and Garbers DL: The primary structure of a plasma membrane guanylate cyclase demonstrates diversity within this new receptor family. Cell 1155–1162, 1989

  15. Pandey KN and Singh S: Molecular cloning and expression of murine guanylate cyclase/atrial natriuretic factor receptor cDNA. J Biol Chem 265: 12342–12348, 1990

    Google Scholar 

  16. Garbers DL: Guanylyl cyclase receptors and their endocrine and autocrine ligands. Cell 71: 1–4, 1992

    Google Scholar 

  17. Fuller F, Porter JG, Arfstein AE, Miller J, Schilling JW, Scarborough RM, Lewicki JA and Schenk DB: Atrial natriuretic peptide clearance receptor: complete sequence and functional expression of cDNA clones. J Biol Chem 263: 9396–9401, 1988

    Google Scholar 

  18. Berl T, Mansour J and Teitelbaum I: ANP stimulates phospholipase C in cultured RIMCD cells: roles of protein kinase C and G protein. Amer J Physiol 260: F590-F595, 1991

    Google Scholar 

  19. Hirata M, Chang C-H and Murad F: Stimulatory effect of atrial natriuretic factor on phosphoinositide hydrolysis in cultured bovine aoratic smooth muscle cells. Biochem Biophys Acta 1010: 346–351, 1989

    Google Scholar 

  20. Anand-Srivastava MB, Srivastava AK and Cantin M: Pertussis toxin attenuates atrial natriuretic factor-mediated inhibition of adenylate cyclase: involvement of inhibitory guanine nucleotide regulatory protein. J Biol Chem 262: 4931–4934, 1987

    Google Scholar 

  21. Anand-Srivastava MB and Trachte GJ: Atrial natriuretic factor receptors and signal transduction mechanisms. Pharmacol Rev 45: 455–489, 1993

    Google Scholar 

  22. Glossman H, Baukal AJ and Catt KJ: Properties of angiotensin II receptors in the bovine and rat adrenal cortex. J Biol Chem 249: 825–834, 1974

    Google Scholar 

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

    Google Scholar 

  24. Girard PR, Mazzei GJ, Wood JG and Kuo JF: Polyclomal antibodies to phospholipid/Ca2+-dependent protein kinase and immunocytochemical localization of the enzyme in rat brain. Proc Natl Acad Sci USA 82: 3030–3034

  25. Towbin H, Staehlin T and Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354, 1985

    Google Scholar 

  26. Brugge JS and Erikson RL: Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature (London) 269: 346–348, 1977

    Google Scholar 

  27. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227: 680–685, 1970

    Google Scholar 

  28. Huang K-P, Chan KFJ, Singh TJ, Nakabayashi H and Huang FL: Autophosphorylation of rat brain Ca2+-activated and phospholipid-dependent protein Kinase. J Biol Chem 261: 12134–12140, 1986

    Google Scholar 

  29. Mochly-Rosen D and Kosland Jr. DE: Domain structure and phosphorylation of protein kinase C. J Biol Chem 262: 2291–2297, 1987

    Google Scholar 

  30. Lee MH and Bell RM: The lipid binding, regulatory domain of protein kinase C: a 32-kDa fragment contains the calcium and phosphatidylserine-dependent phorbol diester binding activity. J Biol Chem 261: 14867–14870, 1986

    Google Scholar 

  31. Melloni E, Pontremali S, Michetti M, Sacco O, Sparatore B and Horecker BL: The involvement of calpain in the activation of protein kinase C in neutrophil stimulated by phorbol myristic acid. J Biol Chem 261: 4101–4105, 1986

    Google Scholar 

  32. Elliott ME and Goodfriend TL: Inhibition of aldosterone synthesis by atrial natriuretic factor. Fed Pro 45: 2376–2381, 1986

    Google Scholar 

  33. Jaiswal N, Paul AK, Jaiswal RK and Sharma RK: Atrial natriuretic factor regulation of cyclic GMP levels and steroidogenesis in isolated fasciculata cells of rat adrenal cortex. FEBS Lett 199: 121–124, 1986

    Google Scholar 

  34. Newton AC and Koshland Jr. DE: Protein Kinase C autophosphorylates by an intrapeptide reaction. J Biol Chem 262: 10185–10188, 1987

    Google Scholar 

  35. Hassid A: Atriopeptin II decreases cytosolic free Ca2+ in cultured vascular smooth muscle cells. Am J Physio 251: C681-C686, 1986

    Google Scholar 

  36. Nambi P, Whiteman M, Aiyer N, Stassen F and Crooke S: Inhibition of formation of cyclic AMP and cyclic GMP by vasopressin in smooth muscle cells is insensitive to pertussis toxin. Biochem J 244: 481–484, 1987

    Google Scholar 

  37. Jaiswal RK, Jaiswal N and Sharma RK: Negative regulation of atrial natriuretic factor receptor-coupled membrane guanylate cyclase by phorbol ester: potential protein kinase C regulation of cGMP signal in isolated adrenocortical carcinoma cells of rat. FEBS Lett 227: 47–50, 1988

    Google Scholar 

  38. Elliott ME and Goodfriend TL: Atrial natriuretic peptide inhibits protein phosphorylation stimulated by angiotensin II in bovine adrenal glomerulosa cells. Biochem Biophys Res Commun 140: 814–820, 1986

    Google Scholar 

  39. Sen I and Roy P: Atrial natriuretic factor-induced phosphorylation of human placental membrane protein: an effect mimicked by guanosine 3′–5′-cyclic monophosphate. Biochem Biophys Res Commun 129: 431–438, 1986

    Google Scholar 

  40. Silver PJ, Kocmund SM, Pinto PB: Enhanced phosphorylation of arterial particulate proteins by cyclic nucleotide and human atrial natriuretic factor. Eur J Pharmacol 122: 385–386, 1986

    Google Scholar 

  41. Tamm C, Lang U and Vallottan MB: Atrial natriuretic factor inhibits angiotensin II and phorbol ester causes proteolytic activation of Ca2+-activated phospholipid-dependent protein kinase. Eur J Biochem 151: 419–423, 1985

    Google Scholar 

  42. Pandey KN: Stimulation of protein phosphorylation by atrial natriuretic factor in plasma membranes of bovine adrenal cortical cells. Biochem Biophys Res Commun 163: 988–994, 1989

    Google Scholar 

  43. Pandey KN, Inagami T, Girard PR, Kuo JF and Misano KS: New signal transduction mechanisms of atrial natriuretic factor: inhibition of phosphorylation of protein kinase C and a 240 kDa protein in adrenal cortical plasma membranes by cGMP-dependent and-independent mechanisms. Biochem Biophys Res Commun 148: 689–695, 1987

    Google Scholar 

  44. Pandey KN, and De S: Regulation of inositol phosphate production by atrial natriuretic factor and 8-bromo-cGMP in bovine iris sphincter smooth muscle. Am Soc Hypert 6: 56A, 1993

    Google Scholar 

  45. Coussens L, Parker PJ, Rhee L, Yang-Feng TL, Chen E, Waterfield MD, Francke U and Ullrich A: Multiple distinct forms of bovine and human protein kinase C suggest diversity in cellular signalling pathways. Science 233: 859–866, 1986

    Google Scholar 

  46. Makowske M, Ballester R, Cayres Y, and Rosen OM: Immunochemical evidence that three protein kinase C isozymes increase in abundance during HL-60 differentiation induced by dimethyl sulfoxide and retinoic acid. J Biol Chem 263: 3402–3410, 1988

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pandey, K.N. Atrial natriuretic factor inhibits autophosphorylation of protein kinase C and A 240-kDa protein in plasma membranes of bovine adrenal glomerulose cells: Involvement of cGMP-dependent and independent signal transduction mechanisms. Mol Cell Biochem 141, 103–111 (1994). https://doi.org/10.1007/BF00926173

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00926173

Key words

Navigation