N-ethoxycarbonyl-D-phenylalanyl-L-prolyl-α-azalysine p-nitrophenyl ester: A novel, high selective and optimal chromogenic active site titrant for human and bovine α-, β- and γ-thrombin

Gianni Balliano, Paola Milla, Cesare Giordano, Carlo Gallina, Massimo Coletta, Enea Menegatti, Menico Rizzi, Martino Bolognesi, Paolo Ascenzi

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)557-561
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume225
Issue number2
DOIs
Publication statusPublished - Aug 14 1996

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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