Method for colorimetric determination of cPNMP specific enzyme activity via coupled enzymatic reactions

摘要

The invention relates to a method for determination of phosphodiesterase enzyme (PDE) activity or the inhibitory activity of an inhibitor candidate molecule by colorimetric assay. The invention primarily relates to the coupled usage of enzymes and enzyme specific substrates for the colorimetric determination of the enzyme activity in the enzyme catalyzed conversion of cyclic purine nucleoside monophosphate (cPNMP) to opened PNMP (e.g. cGMP to GMP). The invention is based on the discovery that four or five enzymes can be coupled in a one pot reaction to provide a final colored mixture that is suitable for qualitative or quantitative analysis.