Agar plate screening procedure for cyclic adenosine 3',5'-monophosphate and inhibitors of cyclic nucleotide phosphodiesterase.
PMID 201216 · Applied and environmental microbiology · 1977 · DOI 10.1128/aem.34.5.604-606.1977
Authors: P J Somers; C E Higgens
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.
Adenylyl Cyclases; Agar; Bacteriological Techniques; Caffeine; Cyclic AMP; Escherichia coli; Fermentation; Galactose; Mutation; Papaverine; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Theophylline
Cite
P J Somers; C E Higgens. Agar plate screening procedure for cyclic adenosine 3',5'-monophosphate and inhibitors of cyclic nucleotide phosphodiesterase. Applied and environmental microbiology (1977). PMID 201216. DOI 10.1128/aem.34.5.604-606.1977.