Assay of human platelet guanylate cyclase activity by high-performance liquid chromatography with fluorescence derivatization.
A selective and sensitive high-performance liquid chromatography (HPLC) method with fluorescence derivatization for the assay of guanylate cyclase (GC) activity is described. GTP and cGMP, which are the substrate and the product of GC, respectively, and other guanine-containing compounds are selectively converted by the reaction with (3,4-dimethoxyphenyl)glyoxal to the fluorescent derivatives. The derivatives were separated by reversed-phase HPLC. The limit of detection at a signal-to-noise ratio of 3 for cGMP was 10 fmol on the column. The sensitivity of this method was less than that of the conventional radioisotopic method, but this method is simple and convenient. Human platelet GC activity was measured, and the effects of some compounds were investigated.
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
K
K
Soda
Y
Y
Ohba
K
K
Zaitsu
D001792:Blood Platelets / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006162:Guanylate Cyclase / Q000097:blood*; D006801:Humans; D007700:Kinetics; D013050:Spectrometry, Fluorescence
D006162:Guanylate Cyclase