![Preparing and Running a Docking job
A. Preparing the protein
1. Opening file: [Right-click "PMV molecules"] → [choose file].
2. Color by atom: [Click ◊ under "Atom"].
3. Eliminate water: Select → Select from string → [write HOH* in "Residue"
line and * in the "Atom" line] → Add → Dismiss → Edit → Delete → Delete
AtomSet.
4. Find missing atom and repairing them: File → Load module → [Pmv;
repairCommands] → Edit → Misc. → Check for missing atoms → Edit →
Misc. → Repair missing atoms.
5. Add hydrogens: Edit → Hydrogens → Add → [choose "All hydrogens", "no
bond order", and "Yes" to renumbering].
6. Hide protein: [Click on the gray under "showMolecules"].
(Note: if you are planning rigid docking (i.e. no flexible parts in the protein), save the
protein as RH.pdb for now)
B. Preparing the ligand
1. Make sure the ligand has all hydrogens added before working with ADT.
2. [Toggle the "AutoDock Tools" button].
3. Opening file: Ligand → Input → Open → All Files → [choose file] → Open.
(ADT now automatically computes Gasteiger charges, merges nonpolar
hydrogens, and assigns Autodock Type to each atom.)](https://image.slidesharecdn.com/autodock4-procedure-100223012604-phpapp01/85/Autodock4-Procedure-4-320.jpg)
![(Note: if there are problems with the automatic charge assignment on any residue, this
could be addressed by: Edit → Charges → Check Totals on residues. The molecule
is now shown with all the assigned charges, and they could be changed manually.
Alternatively, the deficit charge can be spread over the entire residue.)
4. Define torsions: * Ligand → Torsion Tree → Detect Root (this is the rigid part
of the ligand) * Ligand → Torsion Tree → Choose Torsions → [either
choose from the viewer specific bonds, or use the widget to make certain
bond types active (rotatable) or inactive (non-rotatable). Amide bonds should
NOT be active (colored pink)] → Done.
* Ligand → Torsion Tree → Set Number of Torsions →
[choose the number of rotatable bonds that move the 'fewest' or 'most' atoms].
5. Save ligand file: * Ligand → Output → Save as PDBQT → [save with
L.pdbqt].
6. Hide the ligand, as explained in (A5) for the protein.
C. Preparing the flexible residue file
(Note: if you are planning rigid docking, ignore this section and do the following:
Grid → Macromolecule → Open → [choose RH.pdb]. AutoDock will
automatically add charges and merge hydrogens. Save the object as RH.pdbqt and
move to section D.)
1. Flexible residues → Input → Choose molecule → [choose the original protein
R.pdb] → Yes to merge nonpolar hydrogens (AutoDock assigns charges +
atom types to R.pdb, and merges nonpolar hydrogens).](https://image.slidesharecdn.com/autodock4-procedure-100223012604-phpapp01/85/Autodock4-Procedure-5-320.jpg)
![2. Select the residues to be flexible: Select → Select from string → ARG8 → Add
→ Dismiss.
3. Define the rotatable bonds: Flexible residues → Choose torsions in currently
selected residues → [click on rotatable bonds to inactivate them, or vice
versa].
4. Save the flexible residues: Flexible residues → Output → Save flexible
PDBQT → [save as R_flex.pdbqt].
5. Save the rigid residues: Flexible residues → Output → Save rigid PDBQT →
[save as R_rigid.pdbqt].
6. Delete this version of protein: Edit → Delete → Delete Molecule → [choose
protein (R)] → Delete → Dismiss.
D. Running AutoGrid calculation
The purpose of this section is to define the search grid and produce grid maps
used later by Autodock.
1. Open the rigid protein: Grid → Macromolecule → Open → [choose the rigid
protein] → Yes to preserving the existing charges.
(Note: if you are doing rigid docking, choose RH.pdbqt)
2. Prepare grid parameter file: Grid → Set Map Types → Choose Ligand →
[choose the ligand already opened] → Accept.
3. Set grid properties: Grid → Grid Box → [Set the grid dimensions, spacing,
and center] → File → Close Saving Current.](https://image.slidesharecdn.com/autodock4-procedure-100223012604-phpapp01/85/Autodock4-Procedure-6-320.jpg)
![4. Save the grid settings as GPf file: Grid → Output → Save GPF → [save as
R.gpf].
5. [Make sure the AutoGrid executable is in the same directory as the input
files].
6. Running: Run → Run AutoGrid → [make sure the program name has the
right path, and that it is where the input files are] → Launch → [in the
command prompt prompt, type "tail –f hsg1.glg" to follow the process]
(Note: the AutoGrid calculation can be started directly from the command prompt
by typing "autogrid4 –p hsg1.gpf –l hsg1.glg &")
E. Preparing the docking parameter file (.dpf)
1. Specifying the rigid molecule: Docking → Macromolecule → Set Rigid
Filename → [choose R_rigid.pdbqt]. (or RH.pdbqt for rigid docking)
2. Specifying the ligand: Docking → Ligand → Choose → [choose L.pdbqt] →
[here you can set the initial location of the ligand] → Accept.
3. Specifying the flexible residues: Docking → Macromolecule → Set flexible
Residues Filename → [choose R_flex.pdbqt].
4. Setting the parameters for the chosen docking method: Docking → Search
Parameters → Genetic Algorithm → [for 1st time, use the short number of
evaluations (250,000), and for other runs choose the medium or long] →
Accept.
5. Setting docking parameters: Docking → Docking Parameters → [choose the
defaults].](https://image.slidesharecdn.com/autodock4-procedure-100223012604-phpapp01/85/Autodock4-Procedure-7-320.jpg)
![6. Specifying the name of the ligand dpf file to be formed, containing the docking
instructions: Docking → Output → Lamarckian GA → [type L.dpf].
7. Confirming the details of docking: Docking → Edit DPF → [make sure the
right ligand pdbqt file name appears after the word "move", and that the
right number of active torsions is specified].
Note: "outlev" specifies the level of output detail in the docking result file, in the
section where the conformations are analyzed for similarity. For LGA docking,
outlev=0 is sufficient.
F. Running AutoDock4
1. [Make sure the AutoDock executable is in the same directory as the
macromolecule, ligand, GPF, DPF and flex files (in case of flexible docking)].
2. Running: Run → Run AutoDock... → Launch.](https://image.slidesharecdn.com/autodock4-procedure-100223012604-phpapp01/85/Autodock4-Procedure-8-320.jpg)
![Viewing Docking Results
A. Reading the docking log file (.dlg)
1. [Toggle the "AutoDock Tools" button].
2. Analyze → Dockings → Open → [choose L.dlg].
3. Analyze → Conformations → Load → [double-click on each conformation to
view it on screen].
B. Visualizing docked conformations
1. Analyze → Conformations → Play... (Note: & allows changing the ligand's
color)](https://image.slidesharecdn.com/autodock4-procedure-100223012604-phpapp01/85/Autodock4-Procedure-10-320.jpg)
Autodock4 Procedure
- 1. AutoDock 4 and ADT: Short Tutorial By Amit Kessel
- 2. Overview Working with AutoDock4 includes 3 steps: 1. Preparation of receptor & ligand files. 2. Calculation of affinity maps by using a 3D grid around the receptor & ligand. 3. Defining the docking parameters and running the docking simulation. The preparation step starts with pdb files of receptor (R.pdb) and ligand (L.pdb), which are added hydrogens and then saved as RH.pdb & LH.pdb. The calculation of affinity maps in the "Grid" section requires the above pdb files to be assigned charges & atom types, and also that the nonpolar hydrogens are merged. This is done automatically by ADT, and the resulting files need to be saved as RH.pdbqt & LH.pdbqt, which is the only format AutoGrid & AutoDock can work with. Calculation of affinity maps is done by AutoGrid, and then docking can be done by AutoDock. The newest docking algorithm is LGA (Lamarckian Genetic Algorithm).
- 3. Files Step Input Output File contains: Preparation of R.pdb R_rigid.pdbqt Rigid part of the receptor receptor and ligand R_flex.pdbqt Flexible part of the receptor files L.pdb L.pdbqt AutoGrid R_rigid.pdbqt R.gpf Grid parameters R.glg Grid log file (not used) R.*.map Atom-specific affinity maps R.maps.fld Grid_data_file R.d.map Desolvation map R.e.map Electrostatic map Preparation for R_rigid.pdbqt L.pdf Docking parameters docking R_flex.pdbqt L.pdbqt Docking R_rigid.pdbqt L.dlg Log + coordinates + energies R_flex.pdbqt L.pdbqt L.pdf R.*.map R.maps.fld R.d.map R.e.map
- 4. Preparing and Running a Docking job A. Preparing the protein 1. Opening file: [Right-click "PMV molecules"] → [choose file]. 2. Color by atom: [Click ◊ under "Atom"]. 3. Eliminate water: Select → Select from string → [write HOH* in "Residue" line and * in the "Atom" line] → Add → Dismiss → Edit → Delete → Delete AtomSet. 4. Find missing atom and repairing them: File → Load module → [Pmv; repairCommands] → Edit → Misc. → Check for missing atoms → Edit → Misc. → Repair missing atoms. 5. Add hydrogens: Edit → Hydrogens → Add → [choose "All hydrogens", "no bond order", and "Yes" to renumbering]. 6. Hide protein: [Click on the gray under "showMolecules"]. (Note: if you are planning rigid docking (i.e. no flexible parts in the protein), save the protein as RH.pdb for now) B. Preparing the ligand 1. Make sure the ligand has all hydrogens added before working with ADT. 2. [Toggle the "AutoDock Tools" button]. 3. Opening file: Ligand → Input → Open → All Files → [choose file] → Open. (ADT now automatically computes Gasteiger charges, merges nonpolar hydrogens, and assigns Autodock Type to each atom.)
- 5. (Note: if there are problems with the automatic charge assignment on any residue, this could be addressed by: Edit → Charges → Check Totals on residues. The molecule is now shown with all the assigned charges, and they could be changed manually. Alternatively, the deficit charge can be spread over the entire residue.) 4. Define torsions: * Ligand → Torsion Tree → Detect Root (this is the rigid part of the ligand) * Ligand → Torsion Tree → Choose Torsions → [either choose from the viewer specific bonds, or use the widget to make certain bond types active (rotatable) or inactive (non-rotatable). Amide bonds should NOT be active (colored pink)] → Done. * Ligand → Torsion Tree → Set Number of Torsions → [choose the number of rotatable bonds that move the 'fewest' or 'most' atoms]. 5. Save ligand file: * Ligand → Output → Save as PDBQT → [save with L.pdbqt]. 6. Hide the ligand, as explained in (A5) for the protein. C. Preparing the flexible residue file (Note: if you are planning rigid docking, ignore this section and do the following: Grid → Macromolecule → Open → [choose RH.pdb]. AutoDock will automatically add charges and merge hydrogens. Save the object as RH.pdbqt and move to section D.) 1. Flexible residues → Input → Choose molecule → [choose the original protein R.pdb] → Yes to merge nonpolar hydrogens (AutoDock assigns charges + atom types to R.pdb, and merges nonpolar hydrogens).
- 6. 2. Select the residues to be flexible: Select → Select from string → ARG8 → Add → Dismiss. 3. Define the rotatable bonds: Flexible residues → Choose torsions in currently selected residues → [click on rotatable bonds to inactivate them, or vice versa]. 4. Save the flexible residues: Flexible residues → Output → Save flexible PDBQT → [save as R_flex.pdbqt]. 5. Save the rigid residues: Flexible residues → Output → Save rigid PDBQT → [save as R_rigid.pdbqt]. 6. Delete this version of protein: Edit → Delete → Delete Molecule → [choose protein (R)] → Delete → Dismiss. D. Running AutoGrid calculation The purpose of this section is to define the search grid and produce grid maps used later by Autodock. 1. Open the rigid protein: Grid → Macromolecule → Open → [choose the rigid protein] → Yes to preserving the existing charges. (Note: if you are doing rigid docking, choose RH.pdbqt) 2. Prepare grid parameter file: Grid → Set Map Types → Choose Ligand → [choose the ligand already opened] → Accept. 3. Set grid properties: Grid → Grid Box → [Set the grid dimensions, spacing, and center] → File → Close Saving Current.
- 7. 4. Save the grid settings as GPf file: Grid → Output → Save GPF → [save as R.gpf]. 5. [Make sure the AutoGrid executable is in the same directory as the input files]. 6. Running: Run → Run AutoGrid → [make sure the program name has the right path, and that it is where the input files are] → Launch → [in the command prompt prompt, type "tail –f hsg1.glg" to follow the process] (Note: the AutoGrid calculation can be started directly from the command prompt by typing "autogrid4 –p hsg1.gpf –l hsg1.glg &") E. Preparing the docking parameter file (.dpf) 1. Specifying the rigid molecule: Docking → Macromolecule → Set Rigid Filename → [choose R_rigid.pdbqt]. (or RH.pdbqt for rigid docking) 2. Specifying the ligand: Docking → Ligand → Choose → [choose L.pdbqt] → [here you can set the initial location of the ligand] → Accept. 3. Specifying the flexible residues: Docking → Macromolecule → Set flexible Residues Filename → [choose R_flex.pdbqt]. 4. Setting the parameters for the chosen docking method: Docking → Search Parameters → Genetic Algorithm → [for 1st time, use the short number of evaluations (250,000), and for other runs choose the medium or long] → Accept. 5. Setting docking parameters: Docking → Docking Parameters → [choose the defaults].
- 8. 6. Specifying the name of the ligand dpf file to be formed, containing the docking instructions: Docking → Output → Lamarckian GA → [type L.dpf]. 7. Confirming the details of docking: Docking → Edit DPF → [make sure the right ligand pdbqt file name appears after the word "move", and that the right number of active torsions is specified]. Note: "outlev" specifies the level of output detail in the docking result file, in the section where the conformations are analyzed for similarity. For LGA docking, outlev=0 is sufficient. F. Running AutoDock4 1. [Make sure the AutoDock executable is in the same directory as the macromolecule, ligand, GPF, DPF and flex files (in case of flexible docking)]. 2. Running: Run → Run AutoDock... → Launch.
- 9. When RH and LH already exist 1. Protein: Grid → Macromolecule → choose RH.pdb → (charges & atom types assigned, nonpolar hydrogen merged) → File → save → write PDBQT → save as RH.pdbqt 2. Ligand: Ligand → Input → Open → All Files → choose LH.pdb → (charges & atom types assigned, nonpolar hydrogen merged) → save as LH.pdbqt 3. Set the rest of the grid parameters & calculate map 4. Setting Docking parameters: Docking → Macromolecule → Set Rigid Filename → choose either RH.pdbqt or RH_rigid.pdbqt → Docking → Ligand → Choose → choose LH.pdbqt → set the rest of the docking parameters. 5. Running docking simulation.
- 10. Viewing Docking Results A. Reading the docking log file (.dlg) 1. [Toggle the "AutoDock Tools" button]. 2. Analyze → Dockings → Open → [choose L.dlg]. 3. Analyze → Conformations → Load → [double-click on each conformation to view it on screen]. B. Visualizing docked conformations 1. Analyze → Conformations → Play... (Note: & allows changing the ligand's color)