Fig. 5: Client binding primes the global dynamics and conformational equilibrium of HtpG.

a Structural view of ps-ns motion changes of methyl groups upon client binding, colored by the values of changes in order parameters, ΔS², which is defined as ΔS² = S²_HtpG-Δ131Δ - S²_HtpG. HtpG is shown in gray, with the bound client protein (Δ131Δ) represented in light pink. The figure was prepared using PDB 8K2T. b Structural view of μs-ms timescale motion changes of methyl groups colored by the values of changes in exchange rates on the μs-ms time scale Δk_ex, which is defined as Δk_ex = k_ex^HtpG-Δ131Δ - k_ex^HtpG. c Methyl-TROSY spectral overlay for representative residues that exhibit resonance split in unliganded HtpG (black) and HtpG-Δ131Δ (pink), annotated with transition states and population ratios. d Population distribution analysis for the second conformational state in AMP-PNP-bound HtpG (gray) and AMP-PNP-bound HtpG-Δ131Δ (green). Data are presented as individual data points with mean ± SEM, based on n = 9 biological replicates for each group (nine representative residues selected; refer to Methods for details). Statistical significance was assessed using a two-sided t-test, with p = 0.0006 as indicated by asterisks. e Population analysis of three split states for L76 (left) and L233 (right) in unliganded HtpG (gray) and HtpG-Δ131Δ (pink), presented as bar graphs.