N-ethoxycarbonyl-D-phenylalanyl-L-prolyl-alpha-azalysine p-nitrophenyl ester: a novel, high selective and optimal chromogenic active site titrant for human and bovine alpha-, beta- and gamma-thrombin.
The serine proteinase catalyzed hydrolysis of N-ethoxycarbonyl-D-phenylalanyl-L-prolyl-alpha-azalysine p- nitrophenyl ester (Eoc-D-Phe-Pro-azaLys-ONp) was investigated at pH 6.2 and 21.0 degrees C. The results are consistent with the minimum three-step catalytic mechanism. The acylation step is rate limiting for human (Lys 77 species) and porcine plasmin, and for bovine beta-trypsin, the deacylation rate being limiting, on the other hand, for human and bovine alpha-, beta- and gamma-thrombin. Moreover the M(r) 33,000 species of human urokinase and the neuraminidase-treated porcine pancreatic beta-kallikrein-B do not catalyze the hydrolysis of the tripeptide. According to the specificity properties of the serine proteinases considered. Eoc-D-Phe- Pro-azaLys-ONp shows the characteristics of a novel, high selective and optimal chromogenic active site titrant for human and bovine alpha-, beta- and gamma-thrombin.
Balliano
G
G
Department of Pharmaceutical Chemistry and Technology, University of Torino, Italy.
Milla
P
P
Giordano
C
C
Gallina
C
C
Coletta
M
M
Menegatti
E
E
Rizzi
M
M
Bolognesi
M
M
Ascenzi
P
P
D000818:Animals; D001665:Binding Sites; D002384:Catalysis; D002417:Cattle; D002863:Chromogenic Compounds / Q000378:metabolism / Q000494:pharmacology*; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D009596:Nitrophenols / Q000378:metabolism; D013379:Substrate Specificity; D013552:Swine; D013917:Thrombin / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism
D002863:Chromogenic Compounds; D009596:Nitrophenols; D014357:Trypsin; D013917:Thrombin; D005341:Fibrinolysin; C024836:4-nitrophenol