Fig. 1

The scheme of PTS and the specific interaction of EIIAGlc/HPr complex（PDB: 1GGR）[38]

Fig. 2

1H-15N HSQC spectra of EIIAGlc/HPr solution (black) and EIIAGlc/HPr E5C (a) or EIIAGlc/HPr E66C (b) (red) were compared under 25 ℃. Experimental observations (red dots) and theoretical calculations (black solid lines) of intermolecular PRE of EIIAGlc and HPr E5C (c) or HPr E66C (d); (e) Structure diagram of HPr probe connection site; (f) Schematic diagram of EIIAGlc structure, where areas with large differences between PRE experimental values and theoretical calculated values are shown in red

Fig. 3

(a) The spatial distributions of HPr in EIIAGlc/HPr E5C, EIIAGlc/HPr E66C complex ensemble structures; (b) The plot of ensemble size against the correlation between the experimental and the theoretical PRE value of the calculated ensemble

Fig. 4

Calculated ensemble structures of EIIAGlc/HPr E5C (a) and EIIAGlc/HPr E66C (b) complexes in dilute buffer. The specific complex of EIIAGlc (yellow) and HPr (blue) are shown as cartoon, and the mass center of HPr molecules of the calculated EIIAGlc/HPr E5C and EIIAGlc/HPr E66C complexes are shown as gray dots. According to the location on the EIIAGlc surface, HPr molecules are divided into three clusters marked using dashed cycles

Fig. 5

(a)~(c) The RMSD plots of three encounter complexes in three parallel trajectories; (d)~(f) The distance plots of three encounter complexes in three parallel trajectories; (g)~(i) The structure change schemes of three different encounter complexes during 60 ns under MD simulations, Ec represents encounter complex (Ec), Sc represents specific complex (Sc)