Fig. 4: Role of interface II connecting BiP and Grp94.
From: Mechanism of client loading from BiP to Grp94 and its disruption by select inhibitors
A Predicted sites of BiPI191 (Hsp70I164) and BiPD187 (Hsp70D160) at interface II (PDB: 7KW7). B Grp94 bulk FRET efficiency under apo conditions. The solid line is fit to a single site binding equation (Kd,app: 0.67 ± 0.07 µM). C Grp94 ATPase rates in the absence and presence of 2 µM BiP NBD constructs. D Grp94 ATP-dependent closure rates in the absence and presence of 1 µM BiP NBD constructs. E FP BiP binding assay of either Grp94 or Grp94 MD to different FITC-labeled NBD constructs under ADP conditions. Lines are fit to a single-site binding equation. Kd values from the fitting: 1.40 ± 0.02 µM (BiP NBD + Grp94), 5.8 ± 1.2 µM (BiP NBD + Grp94 MD), 8.2 ± 1.0 µM (BiPD187A NBD + Grp94), 9.0 ± 1.7 µM (BiPI191A NBD + Grp94), 6.5 ± 0.7 µM (BiPD187A NBD + Grp94 MD). Error bars are the SEM from independent replicate measurements ((B) n = 3, C) n = 9, (D) n = 3 except for BiP NBD* where n = 5, (E) n = 3 except Grp94 MD with FITC-NBD and FITC-D187A NBD where n = 6 and n = 5, respectively. Source data are provided as a Source Data file.
