Abstract
This chapter describes the designs to measure cyclic adenosine monophosphate (AMP) phosphodiesterase and cyclic guanosine monophosphate (GMP) phosphodiesterase by reaction sequences using firefly luciferin and luciferase in the final step. These assay systems are simple, sensitive, inexpensive, and reproducible. Intracellular concentrations of cyclic AMP and cyclic GMP affect a number of biochemical and physiologic processes in the cell. The concentration of cyclic nucleotides can be influenced by activating or inhibiting cyclic nucleotide phosphodiesterases, which catalyze the hydrolysis of the cyclic nucleotides to their 5 ' -monophosphate analogs. The assay procedures for these enzymes are based on the quantitative coupling of the product of the phosphodiesterase reaction to adenosine-5 ' -triphosphate. The concentration of adenosine triphosphate (ATP) in the reaction mixture is determined by means of the firefly luciferin–luciferase reaction.
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