Abstract
A procedure is described for the semiquantitative measurement of cyclic adenosine 3',5'-monophosphate (cAMP) and detection of inhibitors of cAMP phosphodiesterase by an agar plate test. The assay organism was an adenyl cyclase-deficient mutant derived from Escherichia coli HfrH. In the presence of an acid base indicator, acid production from barbohydrate metabolism was observed as a yellow zone around filter paper disks containing cAMP. Since yellow zone formation reflects the presence of cAMP, a phosphodiesterase inhibitor can be detected indirectly by the presence of a yellow zone on assay plates from a reaction mixture of an inhibitor, phosphodiesterase, and cAMP. Three known cyclic nucleotide phosphodiesterase inhibitors were active against beef brain phosphodiesterase in this system.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Chambers D. A., Zubay G. The stimulatory effect of cyclic adenosine 3'5'-monophosphate on DNA-directed synthesis of beta-galactosidase in a cell-free system. Proc Natl Acad Sci U S A. 1969 May;63(1):118–122. doi: 10.1073/pnas.63.1.118. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cheung W. Y. Cyclic 3',5'-nucleotide phosphodiesterase. Preparation of a partially inactive enzyme and its subsequent stimulation by snake venom. Biochim Biophys Acta. 1969 Nov 4;191(2):303–315. doi: 10.1016/0005-2744(69)90249-6. [DOI] [PubMed] [Google Scholar]
- Dalton C., Quinn J. B., Burghardt C. R., Sheppard H. Investigation of the mechanism of action of the lipolytic agent 4-(3,4-dimethoxybenzyl)-2-imidazolidinone (Ro 7-2956). J Pharmacol Exp Ther. 1970 Jun;173(2):270–276. [PubMed] [Google Scholar]
- Davies G. E., Rose F. L., Somerville A. R. New inhibitor of phosphodiesterase with anti-bronchoconstrictor properties. Nat New Biol. 1971 Nov 10;234(45):50–51. doi: 10.1038/newbio234050a0. [DOI] [PubMed] [Google Scholar]
- Eron L., Arditti R., Zubay G., Connaway S., Beckwith J. R. An adenosine 3':5'-cyclic monophosphate-binding protein that acts on the transcription process. Proc Natl Acad Sci U S A. 1971 Jan;68(1):215–218. doi: 10.1073/pnas.68.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pastan I., Perlman R. L. The role of the lac promotor locus in the regulation of beta-galactosidase synthesis by cyclic 3',5'-adenosine monophosphate. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1336–1342. doi: 10.1073/pnas.61.4.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perlman R. L., Pastan I. Pleiotropic deficiency of carbohydrate utilization in an adenyl cyclase deficient mutant of Escherichia coli. Biochem Biophys Res Commun. 1969 Sep 24;37(1):151–157. doi: 10.1016/0006-291x(69)90893-6. [DOI] [PubMed] [Google Scholar]
- Perlman R. L., Pastan I. Regulation of beta-galactosidase synthesis in Escherichia coli by cyclic adenosine 3',5'-monophosphate. J Biol Chem. 1968 Oct 25;243(20):5420–5427. [PubMed] [Google Scholar]
- Varmus H. E., Perlman R. L., Pastan I. Regulation of lac messenger ribonucleic acid synthesis by cyclic adenosine 3',5'-monophosphate and glucose. J Biol Chem. 1970 May 10;245(9):2259–2267. [PubMed] [Google Scholar]
- Yokota T., Gots J. S. Requirement of adenosine 3', 5'-cyclic phosphate for flagella formation in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1970 Aug;103(2):513–516. doi: 10.1128/jb.103.2.513-516.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zubay G., Schwartz D., Beckwith J. Mechanism of activation of catabolite-sensitive genes: a positive control system. Proc Natl Acad Sci U S A. 1970 May;66(1):104–110. doi: 10.1073/pnas.66.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]