This database contains raw electrophysiology data and MD data, organised in two parts: part 1 corresponds to electrophysiology traces and part 2 corresponds to MD files. For detailed information see below. Part 1: electrophysiology data Data separated in two main categories: main text data and SI (only) data. The electrophysiology data is named in the following format: main_FnX_CytK mutant_buffer_substrate_cis_applied potential, where n = figure number and X = panel Data from the main text: Figure 2: --> C) CytK WT + S1 substrate (file names start with main_F2C_CytK WT) --> D) CytK 2E-4D (K128D K155D Q145D S151D) + S1 substrate (main_F2D...) --> E) CytK 2E-4D (K128D K155D Q145D S151D) + tzatziki substrate (main_F2E...) --> F) CytK 2E-4D (K128D K155D Q145D S151D) + mujdei substrate (main_F2F...) Figure 4: --> CytK 2E-4D (K128D K155D Q145D S151D/ 4D) + malE219a substrate (main_F4A...) --> CytK 2E-4D (K128D K155D Q145D S151D/ 4D) + H152A-GBP substrate (main_F4B...) --> CytK 2E-4D (K128D K155D Q145D S151D/ 4D) + W30G-W133L-DHFR substrate (main_F4C...) Supporting information figures general name: SI_Sx_CytK mutant_buffer_substrate_cis_applied potential, where x = number figure from supporting information Figure S4. S1 translocation through the K128D K155D CytK mutant nanopore --> SI_S4... Figure S5. S1 translocation through the K128D K155D Q145D CytK mutant nanopore --> SI_S5... Figure S6. S1 translocation through the K128D K155D T147D CytK mutant nanopore --> SI_S6... Figure S7. Translocation of S1 through the 2E-1D-1Q-Q122D-CytK nanopore --> SI_S7... Figure S8. Translocation of S1 through 2E-4D-CytK nanopores --> see F2D main text (main_F2D...) Figure S9. Tzatziki and the CytK 2E-2D nanopore --> SI_S9... Figure S10. Tzatziki translocation through the K128D Q145D S151D K155D CytK nanopore --> see F2D main text (main_F2E...) Figure S11. Tzatziki translocation through the K128D K155D Q145D CytK nanopore --> SI_S11... Figure S12. Tzatziki translocation through the K128D K155D T147D CytK mutant nanopore --> SI_S12... Figure S14. Translocation of mujdei through 2E-4D-CytK nanopores --> see F2D main text (main_F2F...) Figure S22. Translocation of malE219a through 2E-4D-CytK nanopores in 2 M urea --> see F4A main text (main_F4A...) Figure S23: Translocation GBP H152A through the 2E-4D CytK nanopore in 2.4 M urea --> see F4B main text (main_F4B...) Figure S24. Translocation of W30G-W133L-DHFR through 2E-4D-CytK nanopore in 2.6 M urea --> see F4B main text (main_F4B...) Figure S26. MalE219a translocation through the 2E-4D CytK mutant in 1 M and 1.8 M Gu.HCl --> SI_S26... (1 M GuHCl and 1.8 M GuHCl are included in the file name) Figure S27: WT-CytK tested with the malE219a and malE219aD10ssrA proteins in 1.5 M Gu.HCl --> SI_S27... Original SDS-PAGE gels of the substrates in a powerpoint file Part 2: MD data The data corresponding to the MD simulations is bundled in a zip file named MD_files.zip This file contains the following subfiles linked to main text Figure 3: panel A: - main_F3A_CytK_E2-D2 - main_F3A_CytK_E2-D3 - main_F3A_CytK_E2-D4 - main_F3A_CytK_WT panels C, D and E: - main_F3CDE_CytK_E2-D4_rep1 - main_F3CDE_CytK_E2-D4_rep2 - main_F3CDE_CytK_E2-D4_rep3 and files linked to the supporting information Figure S20: - SI_S20_ABC_CytK_E2-D4_TZA11_rep1 - SI_S20_ABC_CytK_E2-D4_TZA11_rep2 - SI_S20_ABC_CytK_E2-D4_TZA11_rep3