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. 2017 Oct 24;6:e29430. doi: 10.7554/eLife.29430

Figure 2. NMR fingerprints of two main functional conformations of BiP.

The isoleucine region of methyl-TROSY spectra of ADP-bound (left) and ATP-bound (right) BiP* (the full-length ATPase deficient T229G BiP construct, in black) overlaid with the spectra of corresponding nucleotide-bound state of isolated NBD* (the ATPase deficient NBD construct without the interdomain linker, residues 1–413, in red) and isolated SBD (in green). (Grey box, right) Blowup of the representative region of methyl-TROSY spectra of ATP-bound BiP*, showing three non-overlapping peak doublets, used to calculate the populations domain-docked (D) and -undocked (U) conformations. The U/D assignments details can be found in Materials and methods; briefly, FL BiP* peaks (black) that overlapping with peaks from the NBD*(1-413) spectrum (blue), were assigned to the domain-undocked conformation (labeled ‘U’; that is, for the corresponding conformation the isoleucine chemical environment is very similar in the FL protein and isolated NBD); the second peak from each doublet was assigned to the domain-docked conformation (labeled ‘D’, that is, for the corresponding conformation the isoleucine chemical environment in the FL protein is significantly different from the chemical environment in the isolated NBD).

Figure 2.

Figure 2—figure supplement 1. NMR fingerprints of the main steps of the BiP allosteric cycle.

Figure 2—figure supplement 1.

The isoleucine region of methyl-TROSY spectra of BiP* (the full-length ATPase deficient T229G BiP construct, in black) overlaid with the spectra of corresponding nucleotide-bound state of NBD* (the ATPase deficient NBD construct without the interdomain linker, residues 1–413, in blue), including ADP-bound, ADP- and P2 (a model substrate peptide HTFPAVL [Marcinowski et al., 2011b])-bound, ATP-bound and ATP- and P2-bound.