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Table 1 Enzymes, substrates and reaction conditions for each proteinase tested

From: ANISERP: a new serpin from the parasite Anisakis simplex

Enzyme [Enzyme] [Substrate] 250 μM Activity buffer
Thrombin 74 nM Boc-Val-Arg -AMC 50 mM Tris–HCl pH 8, 100 mM NaCl
Trypsin 54 nM Boc-Gln-Ala-Arg-AMC 50 mM Tris–HCl pH 8, 1 mM CaCl2, 0.15 M NaCl
Cathepsin G 0.33 nM N-Succinyl-Ala-Ala-Phe-AMC. 50 mM HEPES/NaOH pH 7.5
Cathepsin L 0.83 nM Z-Phe-Arg-AMC Sodium acetate 100 mM pH 5.5, 1 mM EDTA, 4 mM DTT, 0.001% BSA
Plasmine 333 nM Nt-Boc-Val-Leu-Lys-AMC 50 mM Tris–HCl pH 8, 50 mM NaCl
Elastase 1.2 nM N-Methoxysuccinyl-Ala-Ala-Pro-Val-7 amido AMC 25 mM Tris–HCl pH 8, 100 mM NaCl, 1 mM CaCl2
Chymotrypsin 0.012 nM N-succinyl-Lelu-Leu-Val-Tyr-7 amido AMC 100 mM HEPES/ NaOH pH 7.5
Factor Xa 0.054 nM Boc-Ile-Glu-Gly-Arg-7 amido AMC 50 mM Tris–HCl pH 8.3, 5 mM CaCl2, 0.2 mM NaCl
Factor XIa 14,000 nM Boc-Phe-Ser-Arg-7 amido AMC 50 mM Tris–HCl pH 8, 100 mM NaCl, 1 mM CaCl2
Factor XIIa 0.0017 nM Boc-Val-Arg-AMC 4 mM Sodium Acetate-HCl/0.15 M NaCl/pH 5.3