Adenosine 3′,5′-monophosphate phosphodiesterase assay in tissue homogenates

摘要: Abstract 1. A fast, reliable and sensitive method for the assay of 3′,5′-AMP phosphodiesterase (adenosine 3′,5′-monophosphate phosphohydrolase, EC 3.1.4.c) is described, which also applicable to homogenates crude enzyme preparations containing enzymes catalyzing breakdown reaction product 5′-AMP. The has been worked out rat pancreatic homogenate. 2. based on existing methods in 3′,5′-[3H]AMP used as substrate resulting 5′-AMP dephosphorylated a second step by means added 5′-nucleotidase. 3. It shown that already during first incubation mixture radioactive products formed: 5′-AMP, 5′-IMP, 3′,5′-IMP, adenosine, inosine, hypoxanthine adenine. During 5′-IMP are converted into corresponding nucleosides. 4. after applied column anionic exchange resin. All can be eluted with 0.1 M NaHCO3, while unchanged side retained. After liquid scintillation counting eluate, activity calculated.