TY - JOUR
T1 - N-ethoxycarbonyl-D-phenylalanyl-L-prolyl-α-azalysine p-nitrophenyl ester
T2 - A novel, high selective and optimal chromogenic active site titrant for human and bovine α-, β- and γ-thrombin
AU - Balliano, Gianni
AU - Milla, Paola
AU - Giordano, Cesare
AU - Gallina, Carlo
AU - Coletta, Massimo
AU - Menegatti, Enea
AU - Rizzi, Menico
AU - Bolognesi, Martino
AU - Ascenzi, Paolo
PY - 1996/8/14
Y1 - 1996/8/14
N2 - The serine proteinase catalyzed hydrolysis of N-ethoxycarbonyl-D-phenylalanyl-L-prolyl-α-azalysine p-nitrophenyl ester (Eoc-D-Phe-Pro-azaLys-ONp) was investigated at pH 6.2 and 21.0°C. The results are consistent with the minimum three-step catalytic mechanism. The acylation step is rate limiting for human (Lys77 species) and porcine plasmin, and for bovine β-trypsin, the deacylation rate being limiting, on the other hand, for human and bovine α-, β- and γ-thrombin. Moreover, the M(r) 33,000 species of human urokinase and the neuraminidase-treated porcine pancreatic β-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 α-, β- and γ-thrombin.
AB - The serine proteinase catalyzed hydrolysis of N-ethoxycarbonyl-D-phenylalanyl-L-prolyl-α-azalysine p-nitrophenyl ester (Eoc-D-Phe-Pro-azaLys-ONp) was investigated at pH 6.2 and 21.0°C. The results are consistent with the minimum three-step catalytic mechanism. The acylation step is rate limiting for human (Lys77 species) and porcine plasmin, and for bovine β-trypsin, the deacylation rate being limiting, on the other hand, for human and bovine α-, β- and γ-thrombin. Moreover, the M(r) 33,000 species of human urokinase and the neuraminidase-treated porcine pancreatic β-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 α-, β- and γ-thrombin.
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U2 - 10.1006/bbrc.1996.1211
DO - 10.1006/bbrc.1996.1211
M3 - Article
C2 - 8753800
AN - SCOPUS:0030583301
VL - 225
SP - 557
EP - 561
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -