Fig. 5.

Effect of SHK on the conformational dynamics of NEMO/IKKβ. a Comparison of the HDX of IKKβ in the presence and absence of NEMO for the 13 peptides identified from IKKβ over the measured time points (red-1 min; green-5 min; black-10 min) (pink: Increased exchange, blue: decreased exchange). Positive and negative values indicate decreased or increased HDX, respectively, when IKKβ was incubated in the presence of NEMO or alone. Differences > ±7.5% are considered significant. b Comparison of the HDX of IKKβ in the presence of NEMO/SHK and NEMO only for the 13 peptides identified from IKKβ over the measured time points (red-1 min; green-5 min; black-10 min) (pink: Increased exchange, blue: decreased exchange). Differences > ±4.0% are considered significant. c Comparison of the HDX of NEMO in the presence and absence of IKKβ for the 21 peptides identified from NEMO over the measured time points (red-1 min; green-5 min; black-10 min) (pink: Increased exchange, blue: decreased exchange). Positive and negative values indicate decreased or increased HDX, respectively, when NEMO was incubated in the presence of IKKβ or alone. Differences > ±7.5% are considered significant. d Comparison of the HDX of NEMO in the presence of IKKβ/SHK to IKKβ only for the 21 peptides identified from NEMO over the measured time points (red-1 min; green-5 min; black-10 min). (yellow: Increased exchange, blue: decreased exchange). Differences > ±4.0% are considered significant. e Regions showing significant changes of 712–715 (pink) and 735–742 (blue) of IKKβ in dynamics (i.e., HDX) in the presence of SHK compared to IKKβ alone or NEMO/IKKβ are plotted on the crystal structure of the NEMO/IKKβ complex. f Regions showing significant changes of 98–103 (purple) of NEMO in dynamics (i.e., HDX) in the presence of SHK compared to NEMO alone or NEMO/IKKβ are plotted on the crystal structure of the NEMO/IKKβ complex. Regions colored gray indicate insignificant differences in HDX between experimental states or regions not covered by peptides. g Deuterium uptake plots for the peptide located in IKKβ (peptide 735–742, black) in the presence of NEMO (orange), SHK (green), NEMO/SHK (red). h Central binding cavity occupied by SHK (carbon atoms are shown in green and oxygen atoms in red). In this zoom-in view, the surfaces of IKKβ and NEMO are shown in blue and yellow, respectively. i Deuterium uptake plots for the peptide located in NEMO (peptide 94–103, black) in the presence of IKKβ (orange), SHK (green), and IKKβ/SHK (red). j Central binding cavity occupied by SHK (carbon atoms are shown in green and oxygen atoms in red). In this zoom-in view, the surfaces of IKKβ and NEMO are shown in silver and purple, respectively