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. 1971 Apr;68(4):736–739. doi: 10.1073/pnas.68.4.736

A Simple, Sensitive Protein-binding Assay for Guanosine 3′:5′-Monophosphate

Ferid Murad *, Vincent Manganiello †, Martha Vaughan †,‡
PMCID: PMC389031  PMID: 4323786

Abstract

A protein-binding assay for guanosine 3′:5′-monophosphate, which employs an enzyme preparation from lobster tail muscle, is described. The binding constant for cyclic GMP was 2-10 nM for four different enzyme preparations. The method can be set up in several days and permits the detection of as little as 0.5-1 pmol of cyclic GMP. In some instances, it is possible to assay the nucleotide in urine or deproteinized extracts of tissue without purification. For other samples, a simple method for separating and purifying cyclic AMP and cyclic GMP is described.

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Selected References

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

  1. ASHMAN D. F., LIPTON R., MELICOW M. M., PRICE T. D. Isolation of adenosine 3', 5'-monophosphate and guanosine 3', 5'-monophosphate from rat urine. Biochem Biophys Res Commun. 1963 May 22;11:330–334. doi: 10.1016/0006-291x(63)90566-7. [DOI] [PubMed] [Google Scholar]
  2. Appleman M. M., Birnbaumer L., Torres H. N. Factors affecting the activity of muscle glycogen synthetase. 3. The reaction with adenosine triphosphate Mg++, and cyclic 3'5'-adenosine monophosphate. Arch Biochem Biophys. 1966 Sep 26;116(1):39–43. doi: 10.1016/0003-9861(66)90009-9. [DOI] [PubMed] [Google Scholar]
  3. Brooker G., Thomas L. J., Jr, Appleman M. M. The assay of adenosine 3',5'-cyclic monophosphate and guanosine 3',5'-cyclic monophosphate in biological materials by enzymatic radioisotopic displacement. Biochemistry. 1968 Dec;7(12):4177–4181. doi: 10.1021/bi00852a006. [DOI] [PubMed] [Google Scholar]
  4. Gilman A. G. A protein binding assay for adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1970 Sep;67(1):305–312. doi: 10.1073/pnas.67.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Goldberg N. D., Dietz S. B., O'Toole A. G. Cyclic guanosine 3',5'-monophosphate in mammalian tissues and urine. J Biol Chem. 1969 Aug 25;244(16):4458–4466. [PubMed] [Google Scholar]
  6. Hardman J. G., Davis J. W., Sutherland E. W. Effects of some hormonal and other factors on the excretion of guanosine 3',5'-monophosphate and adenosine 3',5'-monophosphate in rat urine. J Biol Chem. 1969 Dec 10;244(23):6354–6362. [PubMed] [Google Scholar]
  7. Hardman J. G., Davis J. W., Sutherland E. W. Measurement of guanosine 3',5'-monophosphate and other cyclic nucleotides. Variations in urinary excretion with hormonal state of the rat. J Biol Chem. 1966 Oct 25;241(20):4812–4815. [PubMed] [Google Scholar]
  8. Kaminsky N. I., Broadus A. E., Hardman J. G., Jones D. J., Jr, Ball J. H., Sutherland E. W., Liddle G. W. Effects of parathyroid hormone on plasma and urinary adenosine 3',5'-monophosphate in man. J Clin Invest. 1970 Dec;49(12):2387–2395. doi: 10.1172/JCI106458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kuo J. F., Greengard P. Cyclic nucleotide-dependent protein kinases. IV. Widespread occurrence of adenosine 3',5'-monophosphate-dependent protein kinase in various tissues and phyla of the animal kingdom. Proc Natl Acad Sci U S A. 1969 Dec;64(4):1349–1355. doi: 10.1073/pnas.64.4.1349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kuo J. F., Greengard P. Cyclic nucleotide-dependent protein kinases. VI. Isolation and partial purification of a protein kinase activated by guanosine 3',5'-monophosphate. J Biol Chem. 1970 May 25;245(10):2493–2498. [PubMed] [Google Scholar]
  11. 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]
  12. Murad F., Manganiello V., Vaughan M. Effects of guanosine 3',5'-monophosphate on glycerol production and accumulation of adenosine 3',5'-monophosphate by fat cells. J Biol Chem. 1970 Jul 10;245(13):3352–3360. [PubMed] [Google Scholar]
  13. Murad F., Rall T. W., Vaughan M. Conditions for the formation, partial purification and assay of an inhibitor of adenosine 3',5'-monophosphate. Biochim Biophys Acta. 1969 Dec 30;192(3):430–445. doi: 10.1016/0304-4165(69)90392-4. [DOI] [PubMed] [Google Scholar]
  14. POSNER J. B., HAMMERMEISTER K. E., BRATVOLD G. E., KREBS E. G. THE ASSAY OF ADENOSINE-3' ,5'-PHOSPHATE IN SKELETAL MUSCLE. Biochemistry. 1964 Aug;3:1040–1044. doi: 10.1021/bi00896a005. [DOI] [PubMed] [Google Scholar]
  15. Walton G. M., Garren L. D. An assay for adenosine 3',5'-cyclic monophosphate based on the association of the nucleotide with a partially purified binding protein. Biochemistry. 1970 Oct 13;9(21):4223–4229. doi: 10.1021/bi00823a026. [DOI] [PubMed] [Google Scholar]

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