Fig. 4: Resonance assignment of the NBD·ATP-bound state.
From: Mechanism of ATP hydrolysis in the Hsp70 BiP nucleotide-binding domain
a Sections of 2D [13C,1H]-methyl-TROSY spectrum of methyl-labeled NBD in presence of the ATP regeneration system (in-cyclo). Sequence-specific resonance assignments of the Ile-[13C1H]δ1 methyl groups in the ADP-bound (light-blue), ADP·Pi-bound (dark-blue) and ATP-bound (red) states are indicated. b Selected sections of 2D [13C,1H]-methyl-TROSY spectra of residue Val50cγ1 in-cyclo (pink) compared to the equilibrium experiments of NBD apo (grey), NBD with 5 mM Pi (green), NBD with 5 mM ADP (light-blue), NBD with 5 mM ADP·Pi (dark-blue) and NBD with 5 mM ATP (yellow). c Methyl groups with significant chemical shift differences between the ATP-bound and ADP·Pi-bound states displayed on the structure of the ATP-bound state (PDB 3LDL). The CSPs are shown in Supplementary Fig. 7a. d Residue-specific populations of the ADP-bound (light-blue), the ADP·Pi-bound (dark-blue) and the ATP-bound state (red). Each bar corresponds to one methyl group. (black line = average populations). e Populations of the ADP-bound, ADP·Pi-bound and ATP-bound states, averaged over all resonances. Data points represent three independent experiments. The bar represents mean and standard deviation. f Average length of the functional cycle of the NBD functional cycle including average state occupancy time.
