[Photometric method of determination of the amidase activity of proteinases using 4-methylcoumaryl-7-amide substrates].
It was shown that 7-amino-4-methylcoumarin (MC-amine), resulted from the enzymatic hydrolysis of 4-methylcoumaryl-7-amide (MC-amide) peptide substrates, may be estimated not only fluorometrically but also photometrically. A photometric method for estimating activity of tissue kallikrein (EC 3.4.21.35) and urokinase (EC 3.4.21.31) is suggested using Z-Phe-Arg-NHMC and Z-Gly-Gly-Arg-NHMC, respectively, as substrates. Kinetic parameters of the enzymatic hydrolysis, as obtained by photometric and fluorometric detection of the MC-amine formed, were in good agreement. The differential coefficient of molar extinction of the substrates and MC-amine at 360 nm was found to be 10,800 M-1 cm-1.
Pozdnev
V F
VF
Rabinovich
S E
SE
Paskhina
T S
TS
D000595:Amino Acid Sequence; D003374:Coumarins / Q000032:analysis*; D004151:Dipeptides; D006801:Humans; D007610:Kallikreins / Q000032:analysis*; D007700:Kinetics; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis*; D010783:Photometry; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis*
D003374:Coumarins; D004151:Dipeptides; D009842:Oligopeptides; C021159:benzyloxycarbonyl-phenylalanylarginine-4-methylcoumaryl-7-amide; C065647:N-benzyloxycarbonylglycyl-glycyl-arginine-4-methylcoumarinyl-7-amide; D007610:Kallikreins; D014568:Urokinase-Type Plasminogen Activator
Fotometricheskiĭ metod opredeleniia amidaznoĭ aktivnosti proteinaz s ispol'zovaniem 4-metilkumaril-7-amidnykh substratov.