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Fig. 1.

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Fig. 1. Deoxypodophyllotoxin (DPT) directly interacts with EGFR or MET. (A) Structure of DPT. (B) Pull down assay. Lysate from HCC827GR cells was incubated with Sepharose 4B-only beads and Sepharose 4B-conjugated DPT and the experiment was administered by described under “Material and methods”. Sepharose 4B beads; lane 1, DPT- Sepharose 4B beads; lane 2, Input control; lane 3, EGFR or MET was immunoprecipitated using DPT-Sepharose 4B beads that DPT binds EGFR or MET ex vivo. (C) Active EGFR (100 ng) or (D) MET (100 ng) was incubated with ATP at various concentrations (0, 10, 100 μM) and DPT-Sepharose 4B or Sepharose 4B. EGFR or MET was bound to DPT competitively with ATP. The pulled-down proteins were confirmed by western blotting. (E) EGFR or (F) MET kinase activity of DPT by ADP-Glo kinase assay. Each experiment was done in triplicate independently, and data represent the mean value ± SD (n = 3). * p < 0.05. (G) A predicted binding pose of DPT for EGFR (left) and MET (right). The receptor for EGFR and MET was available as PDB entry 1M17 and 4XYF, respectively. As shown as the surface representation of the receptor, DPT (stick) was tied in ATP binding and the ligandreceptor interaction was zoomed in details. The transparent sphere representation indicated the hydrophobic sidechain of ATP binding pocket related on the hydrophobic interactions. Notably, the complex of EGFR and MET was stabilized by the hydrophobic interactions.
J. Microbiol. Biotechnol. 2021;31:559~569
© J. Microbiol. Biotechnol.