An optimized continuous assay for cAMP phosphodiesterase and calmodulin.
A continuous spectrophotometric assay for cAMP phosphodiesterase has been optimized and adopted for assaying calmodulin in biological samples. This method utilizes the coupled enzyme reactions of myokinase, pyruvate kinase, and lactic acid dehydrogenase. The effective molar extinction coefficient for this method is 1.25 X 10(4) at 340 nm. A point-assay method capable of handling a large number of samples has also been established. This same procedure can also be adopted for the assay of calcineurin and other calmodulin-binding proteins.
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000032:analysis*; D000818:Animals; D001921:Brain / Q000201:enzymology; D002147:Calmodulin / Q000032:analysis*; D002148:Calmodulin-Binding Proteins; D002417:Cattle; D004285:Dogs; D004593:Electrophorus; D010749:Phosphoprotein Phosphatases / Q000032:analysis; D051381:Rats; D013056:Spectrophotometry, Ultraviolet
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D010749:Phosphoprotein Phosphatases; D015105:3',5'-Cyclic-AMP Phosphodiesterases