Difference between revisions of "User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/01/30"

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(Molecular Fingerprinting and Docking of Compounds to the ADA active site)
(Molecular Fingerprinting and Docking of Compounds to the ADA active site)
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E. Docking
 
E. Docking
 
+
[[Image:Prepwizard2.png|thumb|right|Protein Preparation Wizard_Import and Process Tab]]
 
* Opened the protein databank (PDB) website, www.rcsb.org, and acquired two ADA Bovine structures. The two following structures were chosen by desirable resolutions:
 
* Opened the protein databank (PDB) website, www.rcsb.org, and acquired two ADA Bovine structures. The two following structures were chosen by desirable resolutions:
 
**1KRM (resolution 2.5 angstroms)
 
**1KRM (resolution 2.5 angstroms)
Line 70: Line 70:
 
* On Maestro, opened the aspirin.prj and imported the downloaded structures of ADA.
 
* On Maestro, opened the aspirin.prj and imported the downloaded structures of ADA.
 
* The structures of ADA were protonated according to the pH of 7.4. This was executed from:
 
* The structures of ADA were protonated according to the pH of 7.4. This was executed from:
** Workflows > Protein preparation > add H<sup>+</sup> (see image on the side of the protein preparation wizard dialog box)
+
** Workflows > Protein preparation Wizard
[[Image:Prepwizard2.png|thumb|right|Protein Preparation Wizard]]
+
** On the Import and Process Tab, add H<sup>+</sup> (see image on the right side of the protein preparation wizard dialog box)
 
** Uncheck the delete waters on the dialog box and click preprocess.
 
** Uncheck the delete waters on the dialog box and click preprocess.
 +
** Moving to the Refine Tab, changed the default pH of 7.0 to 7.4 as shown below.
 +
** Then click Optimize
 +
 +
[[Image:Refine75.png]]
 +
  
 +
* After Optimization,
  
  

Revision as of 12:12, 6 February 2013

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Molecular Fingerprinting and Docking of Compounds to the ADA active site

A. Opening Maestro

  • From the assigned computer, a new terminal was created from either of the following steps:

1. Finder > Services > New terminal at folder

-or-

2. press the provided shortcut key F5

  • Then enter $maestro to open the suite.
  • Draw the molecule on the workspace area. Modifications can be made through the Edit panel > build > fragments > atom properties. Sponge the drawn molecule for clarity.
  • To save the molecule, create this entry on the project table -or- from the workspace option > project entry > name and create the project and then save.
  • The next step is to put the molecule under energy minimization to find a stable energetically conformation.


B. Energy Minimization

  • To initiate energy minimization, go to the Applications option > Impact > Minimization.
  • Minimization analyzes the molecule using Classical Physics.
  • A dialog box appears with the force field set on OPLS 2005.
  • Then click the Minimization tab. On the maximum cycles, this was changed to 10,000. The other standard parameters such as gradient criterion (0.01) and energy change criterion (1e-07) were kept. The pH was changed to the desired pH level of 7.4.
  • Knowing there are amino acids present, by changing the pH of the parameters, the structure must also be converted to its alkoxide form at pH 7.4. As a result, through atom types O3 was changed to OM.
  • After all the parameters were set for minimization, the host zorro was chosen and the minimization was started.
  • Double-click the finished minimization on the monitor jobs (from applications option), then label the partial charges of the molecule.
  • Save the project entry.


C. Calculation of Fingerprint

  • Opened Canvas from the Finder by typing $Canvas
  • Created a new project and named it as Aspirin_fingerprint.
  • Imported the energy minimized aspirin saved earlier from maestro.
  • Under applications option, click binary fingerprints > molprint2D
  • Click on the imported molecule and incorporate.
  • Export the molecule by saving it with an extension of .fp
  • A dialog box will appear, choose remove all properties and click ok.
  • Close the project.


D. Screen Fingerprint

  • After closing the previous aspirin project, open the zinc database.
  • Imported the aspirin.fp molecule and allow duplicate mappings.
  • Go to the Applications > similar/distance screen > select aspirin.
    • Tanimoto similarity
  • Selected the fingerprint column and choose the fingerprint (aspirin)
  • Screened the molecules and then incorporate ~ 15,001
  • Clicked the fingerprint screen column and sort in descending order to show molecules closest to aspirin.
  • On the panel, select rows 1-15,001
  • Saved the selection by exporting the project as aspirin_15001.mae along with its properties and click OK.
  • Closed Canvas


E. Docking

Protein Preparation Wizard_Import and Process Tab
  • Opened the protein databank (PDB) website, www.rcsb.org, and acquired two ADA Bovine structures. The two following structures were chosen by desirable resolutions:
    • 1KRM (resolution 2.5 angstroms)
    • 2Z7G (resolution 2.52 angstroms)
  • The pdb.txt of these structures were downloaded.
  • On Maestro, opened the aspirin.prj and imported the downloaded structures of ADA.
  • The structures of ADA were protonated according to the pH of 7.4. This was executed from:
    • Workflows > Protein preparation Wizard
    • On the Import and Process Tab, add H+ (see image on the right side of the protein preparation wizard dialog box)
    • Uncheck the delete waters on the dialog box and click preprocess.
    • Moving to the Refine Tab, changed the default pH of 7.0 to 7.4 as shown below.
    • Then click Optimize

Refine75.png


  • After Optimization,