Mechanistic insights into Zika virus NS3 helicase inhibition by Epigallocatechin-3-gallate: Supplementary Files

Deepak Kumar; Nitin Sharma; Murali Aarthy; Sanjeev Singh; Rajanish Giri

The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/530600 doi: bioRxiv preprint Figure 5 : EGCG inhibits ATPase activity of NS3 helicase. A) Purification of NS3 helicase by Ni-NTA affinity chromatography. A 10% SDS gel was run and stained with Coomassie dye where final his-tagged NS3 helicase fractions were pooled and concentrated by Amicon (10kDa) centrifugal filter. In this figure, Mprotein ladder (Bio-Rad Precision Plus) and L1 contains purified concentrated his-tagged NS3 helicase protein of ZIKV B) Kinetic parameters (Substrate-Velocity curve) calculated for 80 nM NS3 helicase after varying the substrate (ATP) concentration ranging from 50 µM to 2500 µM. C) IC50 calculated for EGCG against NTPase site by incubating 80 nM helicase with varying concentration of EGCG (serially diluted: 1200 nM: 23.40 nM) D) Inhibition constant (Ki) was calculated for 80nM helicase at different concentration of ATP (100, 150, 250, 400, 600 and 1000 µM) and EGCG concentrations were kept at 0.5 and 0.25 µM. author/funder. All rights reserved. No reuse allowed without permission.