Trypsin-like serine proteinase action: determination of the catalytic parameters Ks, k+2 and k+3 under conditions where the substrate exceeds the enzyme concentration

Paolo Ascenzi, Enea Menegatti, Mario Guarneri, Gino Amiconi

Research output: Contribution to journalArticle

Abstract

A method for the determination of the catalytic parameters Ks, k+2 and k+3 describing trypsin-like serine proteinase action has been developed from the quantitative analysis of the kinetics of hydrolysis of two specific chromogenic substrates, N-α-carbobenzoxy-l-arginine p-nitrophenyl ester and N-α-carbobenzoxy-l-lysine p-nitrophenyl ester, catalyzed by porcine pancreatic ß-kallikrein B, bovine ß-trypsin and human urokinase (Mr 54 000 species), under conditions where the concentration of the substrate exceeds that of the enzyme. Values of Ks, k+2 and k+3 have been estimated from the effect of substrate concentration on the apparent first-order rate constant of the time-course of the burst phase of p-nitrophenol release preceding the steady-state reaction.

Original languageEnglish
Pages (from-to)210-214
Number of pages5
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume998
Issue number2
DOIs
Publication statusPublished - Oct 5 1989

Keywords

  • Catalytic mechanism
  • Enzyme mechanism
  • Serine proteinase
  • Trypsin-like enzyme

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Structural Biology
  • Medicine(all)

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