A sensitive assay of human blood platelet cyclic nucleotide phosphodiesterase activity by HPLC using fluorescence derivatization and its application to assessment of cyclic nucleotide phosphodiesterase inhibitors.
A selective and sensitive HPLC measurement of 3',5'-cyclic nucleotide phosphodiesterase (PDE) activity in human platelets using (3,4-dimethoxyphenyl)glyoxal (DMPG) as a fluorogenic reagent for guanine and its nucleosides and nucleotides is described. cGMP, a substrate for PDE, and GMP, which was produced by the enzyme reaction, are selectively converted by the reaction with DMPG to the fluorescent derivatives. The derivatives were separated by reversed-phase HPLC. Human platelet PDE activity was measured and the inhibitory effects of several compounds were investigated.
Ohba
Y
Y
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Soda
K
K
Zaitsu
K
K
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000378:metabolism*; D001792:Blood Platelets / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D005453:Fluorescence; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D012680:Sensitivity and Specificity
D010726:Phosphodiesterase Inhibitors; D015105:3',5'-Cyclic-AMP Phosphodiesterases