Engineering dynamic gates in binding pocket of penicillin G acylase to selectively degrade bacterial signaling molecules

(01-mutants_design.tar.gz) Mutants design:

1. Input structures of ecPGA from the PDB database (PDB IDs: 1GK9, 1GM7, and 1GM9), processed to resemble wild-type state, repaired by RepairPDB module of FoldX 4
2. Double-point mutants preparation, analysis and filtering:
   1. text files including configuration for FoldX 4
   2. inputs and outputs of CAVER 3.02 calculations on FoldX 4 PDB files of ecPGA double-point mutants
   3. input configuration file and TransportTools library 0.9.4 calculations outputs generated based on the inputs produced in step 01 and 02 above
   4. CSV files containing complete information about FoldX 4 stability prediction and geometrical properties from CAVER 3.02 and TransportTools library version 0.9.4 for ecPGA double-point mutants
3. Triple-point mutant preparation, analysis and filtering:
   1. text files including configuration for FoldX 4
   2. inputs and outputs of CAVER 3.02 calculations on FoldX 4 PDB files of ecPGA triple-point mutants
   3. input configuration file and TransportTools library 0.9.4 calculations outputs generated based on the inputs produced in step 01 and 02 above
   4. CSV files containing complete information about FoldX 4 stability prediction and geometrical properties from CAVER 3.02 and TransportTools library version 0.9.4 for ecPGA triple-point mutants

(02-docking.tar.gz) Preparation of protein-ligand complexes using molecular docking for wild-type ecPGA and 6 best designed triple-point mutants with 6 various bacterial signaling molecules:

1. PDB files of ligand, PDBQT files of the receptor and PDB files of the complexes selected from docking experiment:

Full names of presented protein variants:
ecPGA_wt, wild-type Escherichia coli penicillin G acylase
LAF, Phe138αLeu & Met142αAla & Ile177βPhe ecPGA variant internally referred as 1GK9_Repair_22
LSF, Phe138αLeu & Met142αSer & Ile177βPhe ecPGA variant internally referred as 1GK9_Repair_98
MAF, Phe138αMet & Met142αAla & Ile177βPhe ecPGA variant internally referred as 1GK9_Repair_23
MSF, Phe138αMet & Met142αSer & Ile177βPhe ecPGA variant internally referred as 1GK9_Repair_99
VAF, Phe138αVal & Met142αAla & Ile177βPhe ecPGA variant internally referred as 1GK9_Repair_30
YAF, Phe138αTyr & Met142αAla & Ile177βPhe ecPGA variant internally referred as 1GK9_Repair_33
Full names of presented AHLs:
C06, N-hexanoyl-L-homoserine lactone;
C06-3O, N-3-oxo-hexanoyl-L-homoserine lactone;
C08, N-octanoyl-L-homoserine lactone;
C08-3O, N-3-oxo-octanoyl-L-homoserine lactone;
C10, N-decanoyl-L-homoserine lactone;
C12-3O, N-3-oxo-dodecanoyl-L-homoserine lactone

(03-protein_ligand_MDs.tar.gz) Ligand-enzyme complexes molecular dynamics for wild-type ecPGA and 6 best designed triple-point mutants with 6 various bacterial signaling molecules:

1. Force field parameters in Amber format
2. Input coordinates *.inpcrd, parameters *.parm7 and *.pdb files for each complex ready for simulation in Amber
3. Amber input files *.in for minimization, equilibration and production runs
4. Restart files for each stage of the minimization, equilibration and production runs in Amber *.rst format
5. Simulation output files for each stage of the minimization, equilibration and production runs in Amber *.mdout format
6. Output files generated during post-processing of production runs trajectories in a form of text files generated by cpptraj

(04-free_enzymes_MDs.tar.gz) Free enzymes molecular dynamics of 3 best triple-point ecPGA (VAF, YAF and MSF) mutants prioritized based on protein-ligand molecular dynamics simulations and experimental assays:

1. Force field parameters and input coordinates *.inpcrd, parameters *.parm7 and *.pdb files for each complex ready for simulation in Amber format
2. Amber input files *.in for minimization, equilibration and production runs
3. Restart files for each stage of the minimization, equilibration and production runs in Amber *.rst format
4. Simulation output files for each stage of the minimization, equilibration and production runs in Amber *.mdout format
5. Post-processing analysis of generated trajectories:
   1. Text files with distances, CSV files containing result of PCA and clustering, PNG files with clustered PCA results
   2. Inputs and outputs of MDpocket analysis and visualization of the pocket frequency grid as an isomesh
   3. CAVER input configuration files in text format, CAVER output data including parsed CSV and text files for visualization of entrance opening time evolution and cavity profiles inspection
   4. cpptraj generated text files including RMSD, distances and chi1 angles measurements

All plots were generated using matplotlib or seaborn Python libraries. Figures containing structural representations were generated using PyMOL 2.0.1.

(05-ecPGA_VAF_YAF_MSF_penG_MDs.tar.gz) PenG-enzyme complexes molecular dynamics for wild-type ecPGA and 3 best designed triple-point mutants (VAF, YAF, MSF):

1. PenG force field parameters in Amber (GAFF) format
2. Input coordinates *.inpcrd, parameters *.parm7 and *.pdb files for each complex ready for simulation in Amber
3. Amber input files *.in for minimization, equilibration and production runs
4. Restart files for each stage of the minimization, equilibration and production runs in Amber *.rst format
5. Simulation output files for each stage of the minimization, equilibration and production runs in Amber *.mdout format and analysis output files generated during post-processing of production runs trajectories in a form of text files
6. Reactive Stabilization Score [RSS] statistics summarized in CSV files