%0 Figure %A Jang, Sung-Wuk %A Liu, Xia %A Bun Chan, Chi %A A. France, Stefan %A Sayeed, Iqbal %A Tang, Wenxue %A Lin, Xi %A Xiao, Ge %A Andero, Raul %A Chang, Qiang %A J. Ressler, Kerry %A Ye, Keqiang %D 2010 %T Deoxygedunin binds TrkB ECD and elicits its dimerization. %U https://plos.figshare.com/articles/figure/_Deoxygedunin_binds_TrkB_ECD_and_elicits_its_dimerization_/511435 %R 10.1371/journal.pone.0011528.g003 %2 https://ndownloader.figshare.com/files/840995 %K binds %K trkb %K ecd %K elicits %X

(A) [3H]deoxygedunin binds the ECD but not ICD of TrkB receptor. In vitro binding assay with purified TrkB ECD or ICD (10 µg) and [3H]deoxygedunin (upper panel). In vitro binding assay with purified fragments of TrkB ECD (10 µg) and [3H]deoxygedunin. (B) Determination of binding constant. Binding curve (upper panel). Scatchard plot analysis revealed that deoxygedunin binds TrkB with binding constant of 1.4 µM (lower panel). (C) Deoxygedunin provokes TrkB dimerization. mGST-TrkB and HA-TrkB were cotransfected into HEK293 cells, treated with 0.5 µM various gedunins for 30 min. GST-TrkB was pulled down by glutathione beads, the coprecipitated proteins were analyzed with anti-HA HRP antibody. (D) Deoxygedunin induces TrkB autophosphorylation. HEK 293 cells were transfected with various Trk receptors, followed by deoxygedunin treatment for 30 min. Cell lysates were analyzed by immunoblotting assay.

%I PLOS ONE