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Proc Natl Acad Sci U S A. 1970 Sep; 67(1): 305–312.
doi: 10.1073/pnas.67.1.305
PMCID: PMC283204
PMID: 4318781

A Protein Binding Assay for Adenosine 3′:5′-Cyclic Monophosphate

Alfred G. Gilman
Author information Copyright and License information PMC Disclaimer

Abstract

A simple and sensitive assay for adenosine 3′:5′-cyclic monophosphate (cAMP) has been developed that is based on competition for protein binding of the nucleotide, presumably to a cAMP-dependent protein kinase. The nucleotide-protein complex is adsorbed on a cellulose ester filter. Assay conditions are such that a binding constant approaching 10-9 M is obtained, and the assay is thus sensitive to 0.05-0.10 pmol of cAMP.

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Selected References

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

  • Butcher RW, Ho RJ, Meng HC, Sutherland EW. 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]
  • Kakiuchi S, Rall TW. The influence of chemical agents on the accumulation of adenosine 3',5'-Phosphate in slices of rabbit cerebellum. Mol Pharmacol. 1968 Jul;4(4):367–378. [PubMed] [Google Scholar]
  • BRECKENRIDGE BM. THE MEASUREMENT OF CYCLIC ADENYLATE IN TISSUES. Proc Natl Acad Sci U S A. 1964 Dec;52:1580–1586. [PMC free article] [PubMed] [Google Scholar]
  • Goldberg ND, Larner J, Sasko H, O'Toole AG. Enzymic analysis of cyclic 3', 5'-AMP in mammalian tissues and urine. Anal Biochem. 1969 Apr 4;28(1):523–544. [PubMed] [Google Scholar]
  • Aurbach GD, Houston BA. Determination of 3',5'-adenosine monophosphate with a method based on a radioactive phosphate exchange reaction. J Biol Chem. 1968 Nov 25;243(22):5935–5940. [PubMed] [Google Scholar]
  • Brooker G, Thomas LJ, Jr, Appleman MM. 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. [PubMed] [Google Scholar]
  • Steiner AL, Kipnis DM, Utiger R, Parker C. Radioimmunoassay for the measurement of adenosine 3',5'-cyclic phosphate. Proc Natl Acad Sci U S A. 1969 Sep;64(1):367–373. [PMC free article] [PubMed] [Google Scholar]
  • Walsh DA, Perkins JP, Krebs EG. An adenosine 3',5'-monophosphate-dependant protein kinase from rabbit skeletal muscle. J Biol Chem. 1968 Jul 10;243(13):3763–3765. [PubMed] [Google Scholar]
  • Langan TA. Histone phosphorylation: stimulation by adenosine 3',5'-monophosphate. Science. 1968 Nov 1;162(3853):579–580. [PubMed] [Google Scholar]
  • Miyamoto E, Kuo JF, Greengard P. Cyclic nucleotide-dependent protein kinases. 3. Purification and properties of adenosine 3',5'-monophosphate-dependent protein kinase from bovine brain. J Biol Chem. 1969 Dec 10;244(23):6395–6402. [PubMed] [Google Scholar]
  • Kuo JF, 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. [PMC free article] [PubMed] [Google Scholar]
  • POSNER JB, HAMMERMEISTER KE, BRATVOLD GE, KREBS EG. THE ASSAY OF ADENOSINE-3' ,5'-PHOSPHATE IN SKELETAL MUSCLE. Biochemistry. 1964 Aug;3:1040–1044. [PubMed] [Google Scholar]
  • Appleman MM, Birnbaumer L, Torres HN. 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. [PubMed] [Google Scholar]
  • Kuwano M, Schlessinger D. Binding of adenosine 3':5'-cyclic phosphate to G factor of Escherichia coli, and its effects on GTPase, RNase V, and protein synthesis. Proc Natl Acad Sci U S A. 1970 May;66(1):146–152. [PMC free article] [PubMed] [Google Scholar]
  • Gill GN, Garren LD. A cyclic-3',5'-adenosine monophosphate dependent protein kinase from the adrenal cortex: comparison with a cyclic AMP binding protein. Biochem Biophys Res Commun. 1970 May 11;39(3):335–343. [PubMed] [Google Scholar]
  • BUTCHER RW, SUTHERLAND EW. 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]
  • Goldberg ND, Dietz SB, O'Toole AG. Cyclic guanosine 3',5'-monophosphate in mammalian tissues and urine. J Biol Chem. 1969 Aug 25;244(16):4458–4466. [PubMed] [Google Scholar]
  • Ishikawa E, Ishikawa S, Davis JW, Sutherland EW. Determination of guanosine 3',5'-monophosphate in tissues and of guanyl cyclase in rat intestine. J Biol Chem. 1969 Dec 10;244(23):6371–6376. [PubMed] [Google Scholar]
  • Kuo JF, 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]
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