Figure 2
Inhibitory effect of Ltc 1 peptides against dengue NS2B-NS3pro. The recombinant dengue NS2B (G4-T-G4) NS3pro was produced as a recombinant protein in E. coli. (A) The kinetic assay plot for the inhibition of NS2BNS3pro from DENV2 by the Ltc 1 peptide. This assay was performed using increasing concentrations of inhibitor while all other conditions were kept constant. The data were analysed using the Michaelis-Menten model with a nonlinear regression curve fit in Graph Pad Prism (version 5.01) software. The concentrations of peptide were 0, 5, 10, 20, and 40 μM. (B) ELISA binding of Ltc 1 to dengue NS2B-NS3pro. Increasing concentrations of purified dengue NS2B-NS3pro (0, 20, 30 and 50 nM/well) were bound to black 96-well plate with transparent bottom. The Ltc 1 peptide labeled with FITC fluorescence dye (0, 0.1, 0.5, 1, 5, 10, 20, 30, 50 nM) were prepared in were bound to plates for 3 h on ice in dark place. the fluorescence signals of bound Ltc 1 were detected after washing steps using fluorescence spectrophotometer. (C) Determination of the IC50 value of the Ltc 1 peptide at normal physiologic human temperature (37°C). (D) Determination of the IC50 value of Ltc 1 peptide at the temperature of a human with a high fever (40°C).