Fig. 5: Interactions between BiPNBD and GRP94.
From: Conformational plasticity of a BiP–GRP94 chaperone complex
a, Overview of interfaces I and II in the preloading and loading complex conformations. b, Zoomed-in view of interface I of the BiP–GRP94 preloading complex. Interface I of the cytosolic (Hsp70–Hsp90) loading complex is shown for comparison. The arrow points to the GRP94MD β-sheet loop inserted between BiPNBD lobes IA and IIA. Residues in bold were used for mutational analysis. c, Analytical SEC and SDS–PAGE analysis of complex formation between BiP and GRP94 Δ72 interface I mutants. d, Ni-NTA pulldown analysis of His6-tagged BiP and Strep-tagged GRP94 Δ72 interface I mutants and quantification of the pulldown experiment (n = 3 independent replicates; mean ± s.d.). The asterisk indicates BSA in the interaction buffer. e, Zoomed-in view of interface II of the BiP–GRP94 loading complex. Interface II of the cytosolic loading complex is shown for comparison. f, Ni-NTA pulldown analysis of His6-tagged BiP and Strep-tagged GRP94 Δ72-R116G interface II mutant and quantification of the pulldown experiment (n = 3 independent replicates; mean ± sd). The asterisk indicates BSA in the interaction buffer. Protein bands on SDS–PAGE gels were visualized by Coomassie staining.
