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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