Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1968 Nov;47(11):2458–2468. doi: 10.1172/JCI105928

Metabolism of adenosine 3′,5′-monophosphate by epithelial cells of the toad bladder

Paul F Gulyassy 1
PMCID: PMC297411  PMID: 4304652

Abstract

When whole urinary bladders of the toad were incubated with adenosine 3′,5′-monophosphate-3H (cyclic AMP-3H) > 95% of the radioactivity could be extracted from the tissue with trichloroacetic acid (TCA). The TCA-soluble radioactivity was separable by cation-exchange (Dowex-50) chromatography into residual cyclic AMP-3H, 5′-AMP-3H, adenosine diphosphate (ADP)-3H and inosine-3H. Thus, neither substantial tight binding of cyclic AMP to TCA-precipitable cell constituents nor any novel metabolite of cyclic AMP were found.

On exposure of cyclic AMP-3H to a crude homogenate of the epithelial cells scraped from the mucosal face of the bladder, the principal metabolite was inosine-3H, whereas 5′-AMP-3H was either absent or present in undetectible amounts. However, when the homogenate included added 5′-AMP (2 × 10-2 mole/liter), substantial quantities of 5′-AMP-3H were recovered.

Metabolism of cyclic AMP-3H by homogenates of the epithelial cell scrapings from the bladder was strongly stimulated by alkalinization over the range in pH of 6-9. Theophylline inhibited metabolism of cyclic AMP only to a limit of 50%, the inhibition being limited to the OH--stimulated component. These results suggest the possibility that a second pathway for metabolism of cyclic AMP may exist. If such is the case, its relationship, if any, to the ultimate biological effects of cyclic AMP within cells will be studied.

Full text

PDF
2460

Images in this article

Selected References

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

  1. ANDERSON W. A., BROWN E. The influence of arginine vasopressin upon the production of adenosine 3',5'-monophosphate by adenyl cyclase from the didney. Biochim Biophys Acta. 1963 Apr 9;67:674–676. doi: 10.1016/0006-3002(63)91879-1. [DOI] [PubMed] [Google Scholar]
  2. BUTCHER R. W., SUTHERLAND E. W. Adenosine 3',5'-phosphate in biological materials. I. Purification and properties of cyclic 3',5'-nucleotide phosphodiesterase and use of this enzyme to characterize adenosine 3',5'-phosphate in human urine. J Biol Chem. 1962 Apr;237:1244–1250. [PubMed] [Google Scholar]
  3. Butcher R. W., Ho R. J., Meng H. C., Sutherland E. W. Adenosine 3',5'-monophosphate in biological materials. II. The measurement of adenosine 3',5'-monophosphate in tissues and the role of the cyclic nucleotide in the lipolytic response of fat to epinephrine. J Biol Chem. 1965 Nov;240(11):4515–4523. [PubMed] [Google Scholar]
  4. Butcher R. W., Sneyd J. G., Park C. R., Sutherland E. W., Jr Effect of insulin on adenosine 3',5'-monophosphate in the rat epididymal fat pad. J Biol Chem. 1966 Apr 10;241(7):1651–1653. [PubMed] [Google Scholar]
  5. Chase L. R., Aurbach G. D. Parathyroid function and the renal excretion of 3'5'-adenylic acid. Proc Natl Acad Sci U S A. 1967 Aug;58(2):518–525. doi: 10.1073/pnas.58.2.518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cheung W. Y. Inhibition of cyclic nucleotide phosphodiesterase by adenosine 5'triphosphate and inorganic pyrophosphate. Biochem Biophys Res Commun. 1966 Apr 19;23(2):214–219. doi: 10.1016/0006-291x(66)90530-4. [DOI] [PubMed] [Google Scholar]
  7. EDELMAN I. S., PETERSEN M. J., GULYASSY P. F. KINETIC ANALYSIS OF THE ANTIDIURETIC ACTION OF VASOPRESSIN AND ADENOSINE-3',5'-MONOPHOSPHATE. J Clin Invest. 1964 Nov;43:2185–2194. doi: 10.1172/JCI105092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gulyassy P. F., Edelman I. S. Effect of pH and theophylline on uptake, elution, and antidiuretic action of cyclic AMP. Am J Physiol. 1967 Apr;212(4):740–746. doi: 10.1152/ajplegacy.1967.212.4.740. [DOI] [PubMed] [Google Scholar]
  9. Gulyassy P. F., Edelman I. S. Hydrogen-ion dependence of the antidiuretic action of vasopressin, oxytocin and deaminooxytocin. Biochim Biophys Acta. 1965 May 25;102(1):185–197. doi: 10.1016/0926-6585(65)90212-8. [DOI] [PubMed] [Google Scholar]
  10. HANDLER J. S., ORLOFF J. ACTIVATION OF PHOSPHORYLASE IN TOAD BLADDER AND MAMMALIAN KIDNEY BY ANTIDIURETIC HORMONE. Am J Physiol. 1963 Aug;205:298–302. doi: 10.1152/ajplegacy.1963.205.2.298. [DOI] [PubMed] [Google Scholar]
  11. Handler J. S., Butcher R. W., Sutherland E. W., Orloff J. The effect of vasopressin and of theophylline on the concentration of adenosine 3',5'-phosphate in the urinary bladder of the toad. J Biol Chem. 1965 Nov;240(11):4524–4526. [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Nair K. G. Purification and properties of 3',5'-cyclic nucleotide phosphodiesterase from dog heart. Biochemistry. 1966 Jan;5(1):150–157. doi: 10.1021/bi00865a020. [DOI] [PubMed] [Google Scholar]
  14. ORLOFF J., HANDLER J. S. THE CELLULAR MODE OF ACTION OF ANTIDIURETIC HORMONE. Am J Med. 1964 May;36:686–697. doi: 10.1016/0002-9343(64)90180-9. [DOI] [PubMed] [Google Scholar]
  15. ORLOFF J., HANDLER J. S. The similarity of effects of vasopressin, adenosine-3',5'-phosphate (cyclic AMP) and theophylline on the toad bladder. J Clin Invest. 1962 Apr;41:702–709. doi: 10.1172/JCI104528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. RANDERATH K., RANDERATH E. ION-EXCHANGE CHROMATOGRAPHY OF NUCLEOTIDES ON POLY-(ETHYLENEIMINE)-CELLULOSE THIN LAYERS. J Chromatogr. 1964 Oct;16:111–125. doi: 10.1016/s0021-9673(01)82445-6. [DOI] [PubMed] [Google Scholar]
  17. Robison G. A., Butcher R. W., Oye I., Morgan H. E., Sutherland E. W. The effect of epinephrine on adenosine 3', 5'-phosphate levels in the isolated perfused rat heart. Mol Pharmacol. 1965 Sep;1(2):168–177. [PubMed] [Google Scholar]
  18. SUTHERLAND E. W., OYE I., BUTCHER R. W. THE ACTION OF EPINEPHRINE AND THE ROLE OF THE ADENYL CYCLASE SYSTEM IN HORMONE ACTION. Recent Prog Horm Res. 1965;21:623–646. [PubMed] [Google Scholar]
  19. SUTHERLAND E. W., RALL T. W., MENON T. Adenyl cylase. I. Distribution, preparation, and properties. J Biol Chem. 1962 Apr;237:1220–1227. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES