Journal: Insect biochemistry and molecular biology

Article Title: Heparan sulfate/heparin glycosaminoglycan binding alters inhibitory profile and enhances anticoagulant function of conserved Amblyomma americanum tick saliva serpin 19

doi: 10.1016/j.ibmb.2016.11.002

Figure Lengend Snippet: List of proteases and their substrates used in substrate hydrolysis assays. Lowest effective protease concentrations (LEPC) and rAAS19 concentrations that inhibit less than 80% of protease activity in specified concentration (IC <80%) are specified.

Article Snippet: The statistical significance of HSGAG binding on rAAS19 inhibitory function was validated using unpaired t-test with Welch's correction in Prism 6 software (GraphPad Software). table ft1 table-wrap mode="anchored" t5 caption a7 Protease (Company) LEPC IC <80% Substrate (Company) Tryptase from Human Lung (Sigma-Aldrich) 4.94 nM NI a N-α-Benzoyl-DL-Arg-pNA (Sigma-Aldrich) Kallikrein from Porcine Pancreas (Sigma-Aldrich) 19.05 nM 1 μM H-D-Pro-Phe-Arg-pNA×2HCl (Chromogenix) Proteinase 3, Human Neutrophil (Athens Research & Technology) 779.31 nM NI a N-Metoxysuccinyl-Ala-Ala-Pro-Val-pNA (Santa Cruz Biotechnology) Elastase from Porcine Pancreas (Sigma-Aldrich) 6.18 nM NI a N-Succinyl-Ala-Ala-Ala-pNA (Sigma-Aldrich) Recombinant Human HMW Urokinase (Molecular Innovations) 9.26 nM NI a CH3SO2-D-CHG-Gly-Arg-pNA×AcOH (Pentapharm) Human tPA, >85% Single Chain (Molecular Innovations) 4.73 nM NI a CH3SO2-D-CHG-Gly-Arg-pNA×AcOH (Pentapharm) Papain (Sigma-Aldrich) 2.14 μM 1 μM N-Benzoyl-Phe-Val-Arg-pNA×HCl (Sigma-Aldrich) Bovine Factor IXa beta (Enzyme Research Laboratories) 31.44 nM 1 μM CH3SO2-D-CHG-Gly-Arg-pNA×AcOH (Pentapharm) Factor Xa Protease (New England Biolabs) 4.65 nM 40 nM Benzoyl-Ile-Glu(γ-OR)-Gly-Arg-pNA×HCl (Chromogenix) Human Factor XIa (Enzyme Research Laboratories) 3.69 nM 200 nM H-D-Pro-Phe-Arg-pNA×2HCl (Chromogenix) Human Factor alpha-XIIa (Enzyme Research Laboratories) 2.03 nM 1 μM H-D-Pro-Phe-Arg-pNA×2HCl (Chromogenix) Plasmin from Human Plasma (Sigma-Aldrich) 11.63 nM 20 nM H-D-Val-Leu-Lys-pNA×2HCl (Chromogenix) Thrombin from Bovine Plasma (Sigma-Aldrich) 27.1 nM 100 nM H-D-Phe-Pip-Arg-pNA×2HCl (Chromogenix) Trypsin from Bovine Pancreas (Sigma-Aldrich) 21.01 nM 4 nM Benzoyl-Ile-Glu(γ-OR)-Gly-Arg-pNA×HCl (Chromogenix) Open in a separate window a No inhibition List of proteases and their substrates used in substrate hydrolysis assays.

Techniques: Activity Assay, Concentration Assay, Recombinant