Translocation of linearized full-length proteins through an engineered nanopore under opposing electrophoretic force

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