Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Nov;78(11):6903–6906. doi: 10.1073/pnas.78.11.6903

Phosphorylation of the same specific protein during amylase release evoked by beta-adrenergic or cholinergic agonists in rat and mouse parotid glands.

R Jahn, H D Söling
PMCID: PMC349160  PMID: 6171823

Abstract

Stimulation of amylase secretion from the rat parotid gland by beta-adrenergic agonists is associated with a specific phosphorylation of three membrane-bound proteins designated as proteins I, II, and III [Jahn, R., Unger, C. & Söling, H. D. (1980) Eur. J. Biochem. 112, 345-352]. In contrast, stimuliation by carbachol induced significant phosphorylation of only protein I. This phosphorylation was low compared to isoproterenol-induced phosphorylation but corresponded to the smaller enhancement of amylase secretion. The mouse organ, however, is almost equally sensitive to beta-adrenergic and to cholinergic agonists. Incubation of mouse parotid gland slices with either 20 microM isoproterenol or 10 microM carbachol resulted in strong and comparable releases of amylase, which were accompanied by comparable phosphorylations of protein I. Proteins II and III were phosphorylated only in the presence of isoproterenol. Removal of external calcium by ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetate abolished the carbachol-induced release of amylase but not the phosphorylation of protein I. Isoproterenol-induced secretion of amylase and phosphorylation of proteins I, II, and III were not inhibited under these conditions. Amylase release stimulated by the ionophore A-23187 was accompanied by the phosphorylation of protein I. Two-dimensional electrophoresis revealed that the radioactive spot corresponding to protein I was located at the same position after cholinergic and after beta-adrenergic stimulation, indicating that both stimuli led to the phosphorylation of the same membrane-associated protein. These findings strongly support the view that the phosphorylation of protein I is an important step in the sequence of events leading from receptor activation to exocytosis.

Full text

PDF
6903

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ames G. F., Nikaido K. Two-dimensional gel electrophoresis of membrane proteins. Biochemistry. 1976 Feb 10;15(3):616–623. doi: 10.1021/bi00648a026. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Butcher F. R. Calcium and cyclic nucleotides in the regulation of secretion from the rat parotid by autonomic agonists. Adv Cyclic Nucleotide Res. 1978;9:707–721. [PubMed] [Google Scholar]
  4. Butcher F. R., Putney J. W., Jr Regulation of parotid gland function by cyclic nucleotides and calcium. Adv Cyclic Nucleotide Res. 1980;13:215–249. [PubMed] [Google Scholar]
  5. Butcher F. R. The calcium requirement for stimulation of rubidium efflux from isolated rat parotid acinar cells by carbachol. Life Sci. 1979 May 21;24(21):1979–1982. doi: 10.1016/0024-3205(79)90308-4. [DOI] [PubMed] [Google Scholar]
  6. Jahn R., Unger C., Söling H. D. Specific protein phosphorylation during stimulation of amylase secretion by beta-agonists or dibutyryl adenosine 3',5'-monophosphate in the rat parotid gland. Eur J Biochem. 1980 Nov;112(2):345–352. doi: 10.1111/j.1432-1033.1980.tb07211.x. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  9. Putney J. W., Jr Muscarinic, alpha-adrenergic and peptide receptors regulate the same calcium influx sites in the parotid gland. J Physiol. 1977 Jun;268(1):139–149. doi: 10.1113/jphysiol.1977.sp011851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Putney J. W., Jr Stimulus-permeability coupling: role of calcium in the receptor regulation of membrane permeability. Pharmacol Rev. 1978 Jun;30(2):209–245. [PubMed] [Google Scholar]
  11. Putney J. W., Jr, Weiss S. J., Leslie B. A., Marier S. H. Is calcium the final mediator of exocytosis in the rat parotid gland? J Pharmacol Exp Ther. 1977 Oct;203(1):144–155. [PubMed] [Google Scholar]
  12. Rasmussen H., Goodman D. B. Relationships between calcium and cyclic nucleotides in cell activation. Physiol Rev. 1977 Jul;57(3):421–509. doi: 10.1152/physrev.1977.57.3.421. [DOI] [PubMed] [Google Scholar]
  13. Rudolph S. A., Krueger B. K. Endogenous protein phosphorylation and dephosphorylation. Adv Cyclic Nucleotide Res. 1979;10:107–133. [PubMed] [Google Scholar]
  14. Selinger Z., Naim E. The effect of calcium on amylase secretion by rat parotid slices. Biochim Biophys Acta. 1970 Apr 21;203(2):335–337. doi: 10.1016/0005-2736(70)90148-3. [DOI] [PubMed] [Google Scholar]
  15. Shelby H. T., Gross L. P., Lichty P., Gardner J. D. Action of cholecystokinin and cholinergic agents on membrane-bound calcium in dispersed pancreatic acinar cells. J Clin Invest. 1976 Dec;58(6):1482–1493. doi: 10.1172/JCI108605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Stolze H., Schulz I. Effect of atropine, ouabain, antimycin A, and A23187 on "trigger Ca2+ pool" in exocrine pancreas. Am J Physiol. 1980 Apr;238(4):G338–G348. doi: 10.1152/ajpgi.1980.238.4.G338. [DOI] [PubMed] [Google Scholar]
  17. Vreugdenhil A. P., Roukema P. A. Comparison of the secretory processes in the parotid and sublingual glands of the mouse. 1. Regulation of the secretory processes. Biochim Biophys Acta. 1975 Nov 17;413(1):79–94. doi: 10.1016/0005-2736(75)90060-7. [DOI] [PubMed] [Google Scholar]
  18. Watson E. L., Williams J. A., Siegel I. A. Calcium mediation of cholinergic-stimulated amylase release from mouse parotid gland. Am J Physiol. 1979 May;236(5):C233–C237. doi: 10.1152/ajpcell.1979.236.5.C233. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES