[43] Assay of cyclic nucleotide phosphodiesterase using radiolabeled and fluorescent substrates
RL Kincaid, VC Manganiello - Methods in enzymology, 1988 - Elsevier
Publisher Summary This chapter describes the assay developed in laboratory that yields
quantitative recovery of product with very low background. The reaction catalyzed by cyclic
nucleotide phosphodiesterases involves the breaking of a high-energy phosphodiester
bond in a metaldependent, hydrolytic process. For most enzymes, the preferred metal
cofactors are divalent cations such as Mg 2+ or Mn 2+, although other metals, such as Co
2+, Ni 2+, and Zn 2+, can support lower activity; Ca 2+, by itself, is virtually ineffective. The …
quantitative recovery of product with very low background. The reaction catalyzed by cyclic
nucleotide phosphodiesterases involves the breaking of a high-energy phosphodiester
bond in a metaldependent, hydrolytic process. For most enzymes, the preferred metal
cofactors are divalent cations such as Mg 2+ or Mn 2+, although other metals, such as Co
2+, Ni 2+, and Zn 2+, can support lower activity; Ca 2+, by itself, is virtually ineffective. The …