CHEMBL3887729

Enzyme Assay: Human aminopeptidase P2 (APP2) enzyme activity was determined using the internally quenched fluorescent substrate: H-Lys(2-aminobenzoyl)-Pro-Pro-p-nitroanilide (Bachem, Torrance, Calif.) at 1 μM (=Km) for inhibitor studies and 5 μM for enzyme purification assays. The assay buffer was 0.1 M HEPES, pH 7.4, and the assay temperature was 30° C. Cleavage of the substrate by the enzyme produces an increase in fluorescence due to the release of H-Lys(2-aminobenzoyl). Enzyme rates were determined in an F. fluorescence plate reader (Molecular Devices, Sunnyvale, Calif.) using an excitation wavelength of 320 nm and an emission wavelength of 405 nm.