Analysis of cyclic nucleotide phosphodiesterase by isoelectric focusing coupled to a specific activity stain.
The procedure described allowed a convenient analysis of cyclic nucleotide phosphodiesterase. The different phosphodiesterase forms present in a crude cytosolic preparation from rat heart were separated by isoelectric focusing on a polyacrylamide gel plate. Phosphodiesterase activity bands were rendered evident by a specific staining method. They were then characterized by means of their substrate specificity and their sensitivity to selective phosphodiesterase inhibitors. The correspondence between the stain bands and the previously described activity peaks, obtained by a preparative technique and detected by radioisotopic enzyme assay, was also investigated.
D000818:Animals; D003600:Cytosol / Q000201:enzymology; D007525:Isoelectric Focusing; D008297:Male; D009206:Myocardium / Q000201:enzymology; D010726:Phosphodiesterase Inhibitors; D010727:Phosphoric Diester Hydrolases / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling; D013379:Substrate Specificity
D010726:Phosphodiesterase Inhibitors; D010727:Phosphoric Diester Hydrolases