Cyclic Nucleotide Phosphodiesterase Assay Technology

Because of their critical role in modulating cellular cyclic nucleotide levels, phosphodiesterases (PDEs) are involved in many disease-related signaling pathways. The PDE family is large and diverse, with members having different tissue distribution, sub-cellular localizations, and substrate specificities. Because of these characteristics, the PDEs represent a broad group of potential drug targets. Described in the present unit are the assay development and validation procedures needed to establish a high-throughput screening system for these important enzymes. The assays provide a structured approach for determining the kinetic parameters of related enzyme families to facilitate the characterization of PDE inhibitors. Curr. Protoc. Pharmacol. 49:3.12.1-3.12.26. © 2010 by John Wiley & Sons, Inc. Keywords: phosphodiesterase; PDE assay technologies; PDE assay protocols