Science Advances

Supplementary Materials

The PDF file includes:

  • Fig. S1. Assignments of back-exchanged deuterated NaK2K in the presence of 50 mM K+.
  • Fig. S2. 2D 15N-13Cα correlation spectrum of uniformly (13C and 15N)-labeled NaK2K in the presence of 50 mM K+.
  • Fig. S3. Predicted H/D exchange pattern based on MD simulations.
  • Fig. S4. Occupancies of the main ion binding sites of the NaK2K SF in MD simulations performed without an applied voltage.
  • Fig. S5. Comparison of 2D (H)NH spectra and illustrations of the washing procedure for back-exchanged deuterated NaK2K.
  • Fig. S6. Comparison of 2D (H)NH spectra of back-exchanged deuterated NaK2K in the presence of K+ using through-bond or through-space transfer.
  • Fig. S7. 2D HN slices from 3D H(H)NH spin diffusion experiments with varying mixing times.
  • Fig. S8. Spin diffusion buildup plots from bound water (blue circles) to amide protons and between neighboring amide protons (red triangles) in back-exchanged deuterated NaK2K in the presence of 50 mM K+.
  • Fig. S9. Spin diffusion buildup plots from bound water (blue circles) to amide protons and between neighboring amide protons (red triangles), recorded after the washing procedure for back-exchanged deuterated NaK2K in the presence of 50 mM K+.
  • Fig. S10. The relationship between carbonyl flipping and water presence at S3 in MD simulations under an applied voltage performed with the AMBER force field.
  • Fig. S11. The relationship between carbonyl flipping and water presence at S3 in MD simulations under an applied voltage performed with the CHARMM force field.
  • Table S1. Chemical shift assignments of deuterated back-exchanged NaK2K, in the presence of 50 mM K+, based on 1H-detected solid-state NMR experiments.
  • Table S2. Chemical shift assignments of NaK2K SF residues, in the presence of 50 mM K+, based on 13C-detected solid-state NMR experiments.
  • Table S3. Distances (Å) to amide protons of the SF in NaK2K (PDB ID: 3ouf) from the closest water molecule and binding sites (S1 to S4).
  • Table S4. Experimental parameters for 3D (H)CANH experiments acquired at 25°C on 100% H2O back-exchanged uniformly (2H, 13C, and 15N)-labeled NaK2K.
  • Table S5. Experimental parameters for 3D (H)CONH experiments acquired at 25°C on 100% H2O back-exchanged uniformly (2H, 13C, and 15N)-labeled NaK2K.
  • Table S6. Experimental parameters for 3D (H)CACO(N)H and (H)COCA(N)H experiments acquired at 25°C on 100% H2O back-exchanged uniformly (2H, 13C, and 15N)-labeled NaK2K.
  • Table S7. Experimental parameters for 3D H(H)NH spin diffusion experiments acquired at 25°C on 100% H2O back-exchanged uniformly (2H, 13C, and 15N)-labeled NaK2K.
  • Legend for movie S1

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mp4 format). Typical K+ ion permeation process through NaK2K, observed in a 20-ns trajectory segment of MD simulations under an applied voltage.

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