Agricultural University of Athens - Biotechnology Lab

Homology Modeling 
</COMMENT> NoJDK1.2 support for APPLET!!&nbsp;

Manual

Contents

1. Introduction
2. Program Window
3. User Interaction
    3.1 Menus
    3.2 Keyboard Button Actions
    3.3 Mouse Button Actions
4. Sequence Alignment
5. Internal Residue Calculations
6. Secondary Structure Prediction
7. Least Square Fit - LsqFit module
8. Model Construction - Franky
9. Installation
 
 



1. Introduction                                                                                         Back to Contents

HoMo is a homology modeling program for protein macromolecules. It includes modules for sequence alignment, secondary structure prediction, least square fitting of tertiary structures and construction of models. As input accepts aminoacid sequence files in FASTA and PIR format and PDB files. The output is always the 3D structure of the predicted molecule in PDB format. For the construction of the target molecule multiple protein sequences can be used as templates. The user manipulates the sequence and selects options though a visual editor window in the form of a Java applet.



2. Program Window                                                                                  Back to Contents

The sequences with all the additional information (like secondary structure predictions, internal residues and user selections) are displayed in an applet window.
Internal residues and secondary structure predictions are displayed below each sequence. The residue colors and their relevant information can be customized by the user.
Most of the sequence manipulation and the activation of different molecules is taking place through menu selections. A brief description of them appears at the bottom of the editor window as they get selected.



3. User Interaction                                                                                    Back to Contents

    3.1 Menus                                                                                              Back to Contents
           File Menu : Basic input - output operations
           Edit: Sequence manipulation and selection of program variables
           Display :  Selection of display information
           Run : Execution of modules on the server
           Help : Information in English and Greek

    3.2 Keyboard Button Actions                                                               Back to Contents
            Arrow keys : Move cursor within sequences
           Home : Move to the beginning of sequence
           End : Move to  the end of sequence
           Page Up : Move to the first sequence
           Page Down : move to the last sequence
           Insert : Inserts space at current position
           Delete : Delete space at current position
           Backspace : Delete space to the left of current position and advance cursor to the left
           Shift + Left or Right Arrow : Underline - De underline residues for model construction
           F5 : Select columns as references for least squares fitting
           F9 : Select target sequence. All alignment information is referenced according to the
            target sequence. Reselect to cancel. Same effect can be achieved through menu selection -
            Edit - Target
           #, ~ : Marker symbols. The user can use them to specify residues with special
            characteristics like active center residues.

    3.3 Mouse Button Actions                                                                    Back to Contents
           Left Mouse Button
                Click : Move cursor to pointer position
                Press + Hold + Drag : Underline - De underline residues for model construction
           Middle Mouse Button Click : Inserts space at current position
           Right Mouse Button Click : Delete space at current position



4. Sequence Alignment                                                                             Back to Contents

At present CLUSTALW is executed on the server and the results are presented in the program window.



5. Internal Residue Calculations                                                              Back to Contents

This module calculates internal residues. The molecule space is divided in identical cube cells. The user defines the cube dimension and the module marks with the letter "i"  all the residues that are surrounded along the coordinate axes with filled cells.



6. Secondary Structure Prediction                                                            Back to Contents

The methods of Chou and Fasman, Burgess, Garnier, Lim and Kimura are implemented.



7. Least Square Fit - LsqFit module                                                         Back to Contents

A grid method is used for the 3D alignment. The user selects (by pressing F9) a minimum of 4 columns from the program window. The Ca coordinates of the selected residues are used for the evaluation of the 3D fit based on the least squares criterion.
The geometric center of the selected Ca atoms is used as reference point. The module initially transfers the 3D structures so as to coincide all geometric centers. It continues with rotations and translations covering the conformation space of the possible orientations. It progresses from large steps to smaller until convergence.



8. Model Construction - Franky                                                               Back to Contents

The user selects aminoacid chain fragments (by underlining) from the known structures. Initially a skeleton is constructed by adopting the aminoacid chain coordinates of the selected fragments (from their PDB files). Then the side chains are constructed by adopting the coordinates of the common atoms between the side chain atoms of the known and target proteins at every residue position. The remaining atoms for the completion of the target molecule side chains are added by adopting the most frequently observed conformations. The module also refines the structure by relaxing the bonds of conflicting conformations. At this stage only rotations are performed.



9. Installation                                                                                                Back to Contents

You need Java Plug-in 1.2 and above to view the applet.
Download from  http://java.sun.com/products/plugin/old.html
Also you need to download the policy file  Policy  and copy to:
Solaris     :  your home directory
Windows : C:\WINDOWS
This will allow the applet to read input files (PIR,FASTA,PDB and Work) for your local drives.