josé r. valverde cnb/csic [email protected] · specify net charges for nonprotein molecules in...
TRANSCRIPT
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Autodock tutorial
VINA with UCSF Chimera
José R. ValverdeCNB/CSIC
© José R. Valverde, 2014
CC-BY-NC-SA
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Loading the receptor
Open UCSF Chimera and then load the protein:● File → Open● Documents/epsilon-zeta/1GVN.pdb
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Preparing the receptor
Tools → Surface/Binding Analysis → DockPrep
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Clean Up the structure
Remove solvent and fix nonstandard residues used for crystallization
Add H and charges
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Adding Hydrogens
Note that you can specify the protonation state for specific residues if needed
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Assigning charges
Protein charges are assigned using an AMBER forcefield
AMBER does not contain charges for most nonprotein atoms:
● AM1BCC (semiempirical quantum mechanics, accurate but slow)
● Gasteiger (empirical rules, fast)
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Specify net charges
For nonprotein molecules in the model (if included)
● Specify expected net charge of the whole molecules● Specify how to compute the charges for each atom in
the molecules
Generally (for protein residues and nonprotein molecules) Chimera will check that the sum of atomic charges leads to an integer matching the expected summary charge.
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Wait and save results
Save the “clean” receptor as a “MOL2” file● 1gvn.mol2
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Open the ligand
File → Open → UNAG.mol2
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Prepare the docking job
Tools → Surface/Binding Analysis → AutoDock Vina
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Select molecules and site
Give a “base” name for all jobgenerated files● e.g.: 1gvn+unag
Specify receptor and ligand● Receptor: 1GVN● Ligand: UNAG
Select area to explore: a parallelepiped boundary for the search grid
● Enclose the active site using the mouse● Center: 15 – 35 – 60● Size: 20 – 20 – 20
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Set receptor options
Expand/close the section as needed
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Set ligand options
Expand/close the section as needed
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Select the precision
Click on “Advanced options”● Select the number of binding modes to generate (9)● Select the thoroughness of the search: lower values
will run faster but be less accurate.– Since we cannot wait, we'll use the fastest setting (1)
– You will normally use the slowest one – 8● Select the report threshold (poses not scoring within
these kcal/mol of the best will be discarded)
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Select executable
You can run VINA remotely on a public web service or locally
● The remote server is shared with other users● Local runs require that you have VINA installed
We do not want to overload the server with a tutorial
● Select “Local”● Set the executable to “/usr/bin/vina”
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Check
Ensure that everything is OK:● Receptor is “1GVN”, Ligand is “UNAG”● The green grid box fully encloses the active site● The receptor and ligand options are correct● The precision, threshold and poses requested are
what you want● The correct executable has been chosen
Click on “OK”
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Wait
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Wait
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Inspect results
After it is done, UCSF Chimer will open the “ViewDock” interface
● Tabular list of poses selected● You can change sort order clicking on column
headers● You can compute properties and add them to the
table● You can organize poses:
– Viable– Deleted– Purged
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Compute H-Bonds
Hbonds → Add count to entire receptor
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Choose H-bonds
We only want Hbonds between ligand and receptor● Intermodel
We want to distinguish “inexact” hbonds● Color Hbonds not meeting precise criteria
differently
We do not want intramolecule or intraresidue bonds
We want to see them all● If endpoint atom hidden, show endpoint residue
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Inspect H-bonds
Hbonds contribute significantly to stability You can sort the conformers by Hbonds formed
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Too much clutter?
Bring up the “Model Panel” ● Tools → Model Panel
You can activate/deactivate which models are shown
Play with selections to show only relevant information
Select the reference ligand and give it a special color
Play with UCSF Chimera...
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That was all folks!