Fig. 3: Hydrophobic tagging to establish a BiP–GRP94–substrate complex.
From: Conformational plasticity of a BiP–GRP94 chaperone complex
a, Cartoon model of HT2 misfolding upon covalent conjugation to HyT36. b, Limited proteolysis with trypsin to assess the folding state of HT2 conjugated with HyT36, incubated with the HyT36(-Cl) control or DMSO. c, CD spectroscopy of HT2 conjugated to HyT36 or 6-chlorohexanol. Kinetic measurements at the indicated temperatures are shown. d, Pulldown experiments to study the interaction of His6–BiP with HT2misfolded–HyT36 compared to DMSO control. For quantification, data were normalized to the wild-type BiP elution sample (n = 3 independent replicates; mean ± s.d.). e, Same analysis as in d for GRP94 FL and GRP94 Δ72. f, Sequential pulldown of Strep–GRP94 Δ72 and His6–BiP to isolate a GRP94 Δ72–BiP–HT2misfolded complex. The asterisks in d–f indicate BSA in the interaction buffer. Protein bands on SDS–PAGE gels were visualized by Coomassie staining.
