University of Illinois at Urbana-Champaign

Beckman Institute for Advanced Science and Technology

Theoretical and Computational Biophysics Group
Computational Biophysics Workshop

VMD Molecular Graphics

VMD Developer: John Stone Jordi Cohen

Marcos Sotomayor
Elizabeth Villa
June 2005

A current version of this tutorial is available at

http://www.ks.uiuc.edu/Training/Tutorials/
CONTENTS 2

Contents
1 Basics of VMD 5
1.1 Loading a Molecule . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1.2 Displaying the Protein . . . . . . . . . . . . . . . . . . . . . . . . 6
1.3 Exploring Different Drawing Styles . . . . . . . . . . . . . . . . . 7
1.4 Exploring Different Coloring Methods . . . . . . . . . . . . . . . 9
1.5 Exploring Different Selections . . . . . . . . . . . . . . . . . . . . 9
1.6 Multiple Representations . . . . . . . . . . . . . . . . . . . . . . . 11
1.7 Sequence Viewer Extension . . . . . . . . . . . . . . . . . . . . . 12
1.8 Saving your Work . . . . . . . . . . . . . . . . . . . . . . . . . . 13

2 Multiple Molecules and Scripting 15

2.1 Loading Multiple Molecules . . . . . . . . . . . . . . . . . . . . . 15
2.2 Using the Main Window . . . . . . . . . . . . . . . . . . . . . . . 16
2.3 Tcl Scripting Basics and Tk Console . . . . . . . . . . . . . . . . 17
2.4 The atomselect Command . . . . . . . . . . . . . . . . . . . . . 18
2.5 Aligning Two Molecules . . . . . . . . . . . . . . . . . . . . . . . 21
2.6 Using Color to Display the Deviation . . . . . . . . . . . . . . . . 23

3 Trajectories, Macros and Labels 26

3.1 Loading trajectories . . . . . . . . . . . . . . . . . . . . . . . . . 26
3.2 Macros . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.3 Main Menu Animation Tools . . . . . . . . . . . . . . . . . . . . 31
3.4 Labels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
3.5 An Example Tcl Script: Calculating the RMSD of a trajectory . 36
CONTENTS 3

Introduction
This tutorial introduces new users to VMD and its capabilities. It can also
be used as a refresher course for the occasional VMD user wishing to employ
this program more productively. This tutorial has been designed specifically for
VMD 1.8.3 and should take about 3 hours to complete in its entirety.

The tutorial is subdivided into three separate units of increasing complexity.

The first unit covers the basics of molecular graphics representations and will
introduce everything you need to know to generate nice graphics. The other two
units are targeted toward the scientifically-oriented user and focus on scripting
in VMD. While scripting may be skipped by the non-technical users, we encour-
age everyone to give it a try as it provides some very powerful (and easy to use)
tools that cannot be offered by a simple graphical user interface.

The examples in the tutorial will focus on the study of ubiquitin – a small
protein with interesting properties. Throughout the text, some material will be
presented in separate “boxes”. These boxes include complementary information
to the tutorial, such as information about the biological role of ubiquitin, and
tips or shortcuts for using VMD.

Ubiquitin. This tutorial will focus on the visualization of ubiquitin

with VMD. Ubiquitin is a small protein of 76 amino acids, that is
believed to be present in all eukaryotic cells. It is one of the most
conserved of all eukaryotic proteins (the first 74 amino acids form
a structure that is identical in insects, trout, bovines and human)
and it has been identified in the nucleus, cytoplasm and on the cell-
surface. It’s primary role is in protein degradation, where it acts as
a tag for intracellular proteolysis.
CONTENTS 4

Getting Started
You can find the files for this tutorial in the vmd-tutorial-files directory in
/Desktop/Workshop/vmd-tutorial/. Below you can see in Fig. 1 the files and
directories of vmd-tutorial-files.

Figure 1: Directory structure of vmd-tutorial-files

To start VMD, double-click on the VMD application icon in the Applications

folder.
1 BASICS OF VMD 5

1 Basics of VMD
In this unit you will build a nice image of ubiquitin while becoming accustomed
to basic VMD commands. In addition, you will learn how to look for interesting
structural properties of proteins using VMD.

1.1 Loading a Molecule

Our first step is to load our molecule.

A pdb file, 1UBQ.pdb, that contains
the atom coordinates of ubiquitin is
provided with the tutorial.

1 Choose the File → New Molecule...

menu item Fig. 2(a) in the VMD
Main window. Another window,
the Molecule File Browser (b), will
appear in your screen.

2 Use the Browse... (c) but-

ton to find the file 1UBQ.pdb in
vmd-tutorial-files. Note that
when you select the file, you
will be back in the Molecule File
Browser window. In order to actu- Figure 2: Loading a Molecule.
ally load the file you have to press
Load (d). Do not forget to do this!

Now, ubiquitin is shown in your screen in the OpenGL Display window. You
may close the Molecule File Browser window at any time.

Webpdb. VMD can download a pdb file from the Protein Data
Bank if a network connection is available. Just type the four letter
code of the protein in the File Name text entry of the Molecule File
Browser window and press the Load button. VMD will download it
automatically.

Coordinates file. The file 1UBQ.pdb corresponds to the X-ray struc-

ture of ubiquitin refined at 1.8 Å resolution provided by Senadhi
Vijay-Kumar, Charles E. Bugg and William J. Cook, J. Mol. Biol.
(1987) 194, 531. Note that the protein is surrounded by 58 water
molecules, and that hydrogen atoms are not included.
1 BASICS OF VMD 6

1.2 Displaying the Protein

In order to see the 3D structure of our protein we will use the mouse in multiple
modes.

1 In the OpenGL Display, press the

first (left) mouse button down and
move the mouse. Explore what
happens. This is the rotation
mode of the mouse and allows you
to rotate the molecule around an
axis parallel to the screen Fig.
3(a).
Figure 3: Rotation modes.

2 If you press the second (right) button and repeat the previous step, the
rotation will be done around an axis perpendicular to your screen (b) (For
Mac users, the second button is equivalent to hitting the command key
while pressing the mouse button).

3 In the VMD Main window, look

at the Mouse menu (Fig. 4).
Here, you will be able to switch
the mouse mode from Rotation to
Translation or Scale modes.

4 The Translation mode will allow

you to move the molecule around
the screen while holding the first
(left) button down.
Figure 4: Mouse modes.

5 The Scale mode will allow you to zoom in or out by moving the mouse
horizontally while holding the first (left) button down.

It should be noted that the previous actions performed with the mouse do
not change the actual coordinates of the molecule atoms.

Mouse modes. Note that each mouse mode has its own charac-
teristic cursor and its own shortcut key (r: Rotate, t: Translate, s:
Scale) that could be used instead of the Mouse menu. (Be sure to
have the OpenGL Display window active when using the shortcuts.)
Additional information can be found in the VMD user’s guide.
1 BASICS OF VMD 7

Another useful option is the Mouse → Center menu item. It allows you to
specify the point around which rotations are done.

6 Select the Center menu item and pick one atom at one of the ends of the
protein; The cursor should display a cross.

7 Now, press r, rotate the molecule with the mouse and see how your
molecule moves around the point you have selected.

1.3 Exploring Different Drawing Styles

VMD can display your molecule using a wide variety of drawing styles. Here,
we will explore those that can help you to identify different structures in the
protein.

1 Choose the Graphics → Represen-

tations... menu item. A window
called Graphical Representations
will appear and you will see high-
lighted in yellow Fig. 5(a) the cur-
rent graphical representation used
to display your molecule.

2 In the Draw Style tab (b) we can

change the style (d) and color (c)
of the representation. In this sec-
tion we will focus in the drawing
style (the default is Lines).

3 Each Drawing Method has its own

parameters. For instance, change
the Thickness of the lines by using
the controls on the right bottom
part (e) of the Graphical Repre-
sentations window.

4 Now, choose VDW(van der Waals)

from Drawing Method. Each atom
is now represented by a sphere. In
this way you can see more easily
Figure 5: Graphical Representations
the volumetric distribution of the
window.
protein.

5 To see the arrangements of atoms in the interior of the protein, use the
new controls on the right bottom part of the window (e) to change the
1 BASICS OF VMD 8

Sphere Scale to 0.5 and the Sphere Resolution to 13. Be aware that the
higher the resolution, the slower the display of your molecule will be.

6 Note in Coloring Method → Name, each atom has its own color, i.e: O is
red, N is blue, C is cyan and S is yellow.

7 Press the Default button. This allows you to return to the default prop-
erties of the drawing method.

More representations. Other interesting representations are CPK

and Licorice. In the first one, like in old chemistry ball & stick
kits, each atom is represented by a sphere and each bond is repre-
sented by a cylinder. (Radius and resolution of both the sphere and
the cylinder can be modified independently.) The Licorice drawing
method (widely used) also represents each atom as a sphere and
each bond as a cylinder, but the sphere radius cannot be modified
independently.

The previous representations allow you to see the micromolecular details of

your protein. However, more general structural properties can be seen by using
more abstract drawing methods.

8 Choose the Tube style under Drawing Method and observe the backbone
of your protein. Set the Radius at 0.8.

9 By looking at your protein in the tube mode, can you distinguish how
many helices, β sheets and coils are present in the protein?

The last drawing method we will explore is NewCartoon. It gives a simplified

representation of a protein based in its secondary structure. Helices are drawn
as coiled ribbons, β sheets as solid arrows and all other structures as a tube.
This is probably the most popular drawing method to view the overall architec-
ture of a protein.

10 Choose Drawing Method → NewCartoon.

11 Identify now how many helices, betasheets and coils are present in the
protein.

Structure of ubiquitin. Ubiquitin has three and one half turns of

α-helix (residues 23 to 34, three of them hydrophobic), one short
piece of 310 -helix (residues 56 to 59) and a mixed β sheet with five
strands (residues 1 to 7, 10 to 17, 40 to 45, 48 to 50, and 64 to 72)
and seven reverse turns. VMD uses the program STRIDE to com-
pute the secondary structure according to an heuristic algorithm.
1 BASICS OF VMD 9

Figure 6: Licorice, Tube and NewCartoon representations of Ubiquitin

1.4 Exploring Different Coloring Methods

1 Now, let’s modify the colors of our representation. Choose Coloring Method
→ ResType Fig. 5(c). This allows you to distinguish non-polar residues
(white), basic residues (blue), acidic residues (red) and polar residues
(green).

2 Select Coloring Method → Structure (c) and confirm that the NewCartoon
representation displays colors consistent with secondary structure.

1.5 Exploring Different Selections

Let’s look at different independent (and interesting) parts of our molecule.

1 In the Selected Atoms text entry Fig. 5(f) of the Graphical Representations
window delete the word all, type helix and press the Apply button or
hit the Enter/Return key on your keyboard. (Do this every time you type
something.) VMD will show just the helices present in our molecule.

2 In the Graphical Representations window choose the Selections tab Fig.

7(a). In section Singlewords (b) you will find a list of possible selections
you can type. For instance, try to display β sheets instead of helices by
typing the appropriate word in the Selected Atoms text entry.

Combinations of boolean operators can also be used when writing a selection.

1 BASICS OF VMD 10

3 In order to see the molecule without helices and β sheets, type the follow-
ing in Selected Atoms: (not helix)and(not betasheet)

4 In the section Keyword (c) of the Selections tab (a) you can see properties
that can be used to select parts of a protein with their possible values.
Look at possible values of the Keyword resname (d). Display all the Lysines
and Glycines presents in the protein by typing (resname LYS)or(resname
GLY). Lysines play a fundamental role in the configuration of polyubiquitin
chains.

5 Now, change the current represen-

tation’s Drawing Method to CPK
style and the Coloring Method to
ResID in the Draw Style tab. In
the screen you will be able to see
the different Lysines and Glycines.
How many of each one can you
see?

6 In the Selected Atoms text en-

try type water. Choose Coloring
Method → Name. You should see
the 58 water molecules (in fact
only the oxygens) present in our
system.

7 In order to see which water

molecules are closer to the pro-
tein you can use the command
within. Type water and within
3 of protein. This selects all
the water molecules that are
within a distance of 3 angstroms
of the protein. Figure 7: Graphical Representations
window and the Selections tab.

8 Finally, try typing the following selections in Selected Atoms:

Selection Action
protein Shows the Protein
resid 1 The first residues
(resid 1 76) and (not water) The first and last residues
(resid 23 to 34) and (protein) The α helix
1 BASICS OF VMD 11

All the previous options provide you with a powerful tool to explore different
parts of your protein or molecule.

1.6 Multiple Representations

The button Create Rep Fig. 8(a) in

the Graphical Representations window
allows you to create multiple repre-
sentations. Therefore, you can have
a mixture of different selections with
different styles and colors, all displayed
at the same time.

1 For the current representation, set

the Drawing Method to NewCar-
toon and the Coloring Method to
Structure.

2 In Selected Atoms type protein.

3 Press the Create Rep button (a).

Now, using the menu items of
the Draw Style tab and the Se-
lected Atoms text entry, modify
the new representation in order to
get VDW as the Drawing Method,
ResType as the Coloring Method,
and resname LYS typed in as the
current selection.
Figure 8: Multiple Representations of
Ubiquitin.

4 Repeating the previous procedure, create the following two new represen-
tations:

Drawing Style Coloring Method Selection

CPK Name water
VDW ColorID 1 resid 1 76 and name CA
1 BASICS OF VMD 12

5 Create a final representation by pressing again the Create Rep button.

Select Drawing Method → Surf, the Coloring Method → Molecule and type
protein in the Selected Atoms entry. For this last representation choose
in the Material section (c) the Transparent menu item.

6 Note, that with the mouse, you can select the different representations you
have created and modify each one independently. Also, you can switch
each one on/off by double-clicking on it or delete each one by using the
Delete Rep button (b). At the end of this section, the Graphical Repre-
sentations window should look like Fig. 8.

1.7 Sequence Viewer Extension

When dealing with a protein for the first time, it is very useful to find and
display different amino acids quickly. The sequence viewer extension allows you
to pick and display one or more residues easily.

1 Choose the Extensions → Analysis

→ Sequence Viewer menu item. A
window Fig. 9(a) with a list of the
amino acids (e) and their proper-
ties (b)&(c) will appear in your
screen.

2 With the mouse, click over differ-

ent residues (e) in the list and see
how they are highlighted. In ad-
dition, the highlighted residue will
appear in your OpenGL Display
window in yellow and bond draw-
ing method, so you can visualize
it easily. Use the right button of
the mouse to unselect residues.

3 Using the Zoom controls (f) you

can display the entire list of
residues in the window. This is es-
pecially useful for larger proteins

4 Using the shift key while pressing

the mouse button allows you to
pick multiple residues at the same
time. Look at residues 48, 63, 11
and 29 (e). Figure 9: Sequence window.
1 BASICS OF VMD 13

5 Look at the Graphical Representations window, you should find a new

representation with the residues you have selected using the Sequence
Viewer Extension. As you already did before, you can modify, hide or
delete this representation.

Relevance of Lysines. Polyubiquitin chains can be linked by a pep-

tide bond between C- and N-termini or through linkages involving
lysines 48, 63, 11 or 29 (the ones that you selected using the se-
quence window). Different linked chains have different properties
that are related to the functionality of the chain.

Information about residues is color-coded (d) in columns and obtained from

STRIDE. The B-value column (b) shows the B-value field (temperature factor).
The struct column shows secondary structure (d), where each letter means:

T Turn
E Extended conformation (β sheets)
B Isolated bridge
H Alpha helix
G 3-10 helix
I Pi helix
C Coil

Table 1: Secondary Structure codes used by STRIDE.

1.8 Saving your Work

The image that you have created using VMD can be saved, along with all
representations you have created, as a VMD state. This VMD state contains
all the information needed to start a new VMD session from it, without losing
what you have done.

1 In the VMD Main window, choose the File → Save State menu item. Write
an appropriate name (e.g., myfirststate.vmd) and save it.
The File → Load State menu item will allow you to load a previously saved
VMD state, just like the file you saved. Although the VMD state allows you to
work with the image and explore the properties of our protein using VMD, you
usually need pictures that can be used in articles or other kind of documents.
VMD can render the image you created and generate an image file that can be
used in other applications, as it is shown in the following steps.
1 BASICS OF VMD 14

2 Using all that you have learned until now, find an appropriate view of
the protein by scaling, rotating and translating the molecule. Turn dif-
ferent representations on and off and improve the resolution and different
properties of the selections you have made. If you want an image of high
quality, put special attention to the resolution of each representation.

3 Be aware of the new representations you created with the Sequence Viewer
Extension and hide or delete them if necessary.

4 Before rendering the image, change the background color by choosing the
Graphics → Colors menu item. There, Choose the Display category, the
Background name and the 8 white color. The background should be white
now.

5 Choose the File → Render... menu item. A window called File Render
Controls will appear in your screen.

6 You can render the image using different packages. Pick TachyonInternal
in the Render using menu.

7 Write the name of the file where the image will be saved in the Filename
text entry, i.e., picture.tga (default is plot.tga).

8 Press the Start Rendering button and the file with your image will be
created. Note that this could take some time. You should end up with an
image file named picture.tga (MacOS X or Unix) or picture.bmp (on
Windows).

9 Close the application that opened the image file in order to continue using
VMD.

Now you are done with the first unit of the tutorial. We hope you have learned
the basic commands of VMD. Also, you have generated two files. The first one
is a VMD state that allows you to restart a VMD session, and use or modify all
that you did in this unit. The second file is an image file of your protein that
can be used in other image-viewing applications.
2 MULTIPLE MOLECULES AND SCRIPTING 15

2 Multiple Molecules and Scripting

In the second unit, you will learn to deal with multiple molecules simultaneously.
You will also cover the basics of Tcl scripting by using it to edit atomic data,
align two molecules, and color a molecule according to calculated properties.
1 Start with a new VMD session. If you have just completed unit 1, you
should quit VMD and then launch it again.

2.1 Loading Multiple Molecules

First, you will load the molecules that you will need.

1 Open the Molecule File Browser from the File → New Molecule. . . menu
item.
You need to load the X-ray crystal structure of ubiquitin. You can do this in
the same way you did in Unit 1, or directly in the command line:

2 Make sure you are in the vmd-tutorial-files. In the VMD terminal,

type mol new 1UBQ.pdb.
An equilibration simulation for ubiquitin solvated in a water box has been per-
formed previously for a duration of 1 ns. Your files contain the coordinates of
the last frame of this equilibration. You will now compare the conformation of
ubiquitin at the end of this simulation with the initial crystal structure.

Coordinate files and structure files. To save space, simulation

output files usually contain only the atom coordinates and do not
store unchanging information such as the atom types, atom charge,
segment names and bonds. The latter information is stored in a
separate “structure” file (e.g., a PSF file). To visualize the results
of a simulation, you need to merge both a structure and a coordinate
file into the same molecule.
3 Now, start loading the simulation results into a second molecule. Staying
in the Molecule File Browser, select New Molecule from the menu at the top
of the window. Browse for the file ubiquitin.psf in vmd-tutorial-files,
then click on the Load button. You have just created a new molecule with
a structure but no coordinates.
4 Notice that the menu at the top of the window now says 1: ubiquitin.psf.
This ensures that the next file that you load will be added to that molecule
(molecule ID 1). You do not need to change this menu to New Molecule,
since you are merging data with the PSF file. Now, browse for the
file ubiquitin-equilibrated.coor in vmd-tutorial-files and click on
Load again. This will load the coordinates and merge them with the pre-
viously loaded structure information.
2 MULTIPLE MOLECULES AND SCRIPTING 16

You should now see two superimposed and unaligned molecules. One is the
original crystal ubiquitin. The other one is surrounded by a cube full of water
molecules.

2.2 Using the Main Window

You will now give names to your molecules so that you can identify them later.

1 Double-click on the name of the first molecule in the Main window molecule
listing. The Rename Molecule dialog box should pop up. Type in crystal.
Do the same thing for the second molecule, and call it simulation.
At this point, your Main window should look like Fig. 10. In front of the
molecule name, there are four letters that you can use to manipulate your
molecules.

Figure 10: Main window with initial and final coordinates loaded into two
separate molecules.

F stands for “Fixed,” meaning that that molecule won’t move when you move
the scene around. When the F is black, the molecule is fixed; when it is grayed
out, the molecule is mobile.

2 Double-click on the F to the left of a molecule description in the Main

form. Then, try translating the scene with the mouse while one molecule
is fixed. Do it again with both molecules fixed.
3 When you are done, unfix both molecules and select the Display →
Reset View menu item to correctly reposition the two molecules relative
to each other.
4 Next, double-click on the D of one molecule. The D stands for “Displayed”
and when the D is grayed out it means that the molecule is hidden. You
can control the visibility of all your molecules by double-clicking here.
2 MULTIPLE MOLECULES AND SCRIPTING 17

5 When you are done, make sure that only the crystal molecule is “Dis-
played” and that none of the molecules are “Fixed.”

6 Finally, double-click under the T column to the left of the crystal molecule.
The letter T should now appear in front of it. This makes it the unique
“top” molecule. Making a molecule top makes it a target for scripting
commands.

2.3 Tcl Scripting Basics and Tk Console

VMD includes support for the Tcl/Tk scripting language. This section will
attempt to provide the minimal amount of scripting that you need to know to
run very useful features.

The Tcl/Tk scripting language. Tcl is a rich language that con-

tains many features and commands, in addition to the typical con-
ditional and looping expressions. Tk is an extension to Tcl that
permits the writing of graphical user interfaces with windows and
buttons, etc. More information and docs about the Tcl/Tk language
can be found at http://www.tcl.tk/doc.

To execute Tcl commands, you will be using a convenient text console called
Tk Console.

1 Select the Extensions → Tk Console menu item. A console window should

appear with a prompt (Fig. 11). You can now start entering Tcl/Tk
commands in it.

Figure 11: The Tk Console console.

You will initially focus on the very basic of Tcl/Tk. Here are Tcl’s set and
puts commands:
2 MULTIPLE MOLECULES AND SCRIPTING 18

set variable value – sets the value of variable

puts $variable – prints out the value of variable

2 Try the following commands:

set x 10
puts "the value of x is: $x"
set text "some text"
puts "the value of text is: $text."

As you can see, $variable refers to the value of variable.

Here is a command that performs mathematical operations:

expr expression – evaluates a mathematical expression

3 Try experimenting with the expr command:

expr 3 - 8
set x 10
expr - 3 * $x

One of the most important aspects of Tcl is that you can embed Tcl commands
into others by using brackets. A bracketed expression will automatically be sub-
stituted by the return value of the expression inside the brackets:

[expr.] – represents the result of the expression inside the brackets

4 Create some commands using brackets and test them. Here is an example:
set result [ expr -3 * $x ]
puts $result

2.4 The atomselect Command

You can edit atomic properties using VMD’s atomselect command. The fol-
lowing examples will show you how.

1 After making sure that the crystal molecule is the top molecule (if not,
double-click on T). Open the Graphical Representations window using the
Graphics → Representations. . . menu item.
2 MULTIPLE MOLECULES AND SCRIPTING 19

The PDB B-factor field. The “B” field of a PDB file typically
stores the “temperature factor” for a crystal structure and is read
into VMD’s “Beta” field. Since we are not currently interested in
this information, we can recycle this field to store our own numerical
values. VMD has a “Beta” coloring method, which colors atoms
according to their B-factors. By replacing the Beta values for various
atoms, you can control the color in which they are drawn. This is
very useful when you want to show a property of the system that
you have computed, which is not supported out-of-the-box by VMD.

2 Select the molecule called “crystal” in the Selected Molecule pull-down

menu at the top of the form. Type in protein as the atom selection,
change its Coloring Method to Beta and its Drawing Method to VDW. Your
molecule should now appear as a mostly red and green assembly of spheres.

You will now learn a very important Tcl command in VMD:

atomselect molid selection – creates a new atom selection

The first argument to atomselect is the molecule ID (shown to the very left of
the Main form), the second argument is a textual atom selection like what you
have been using to describe graphical representations in unit 1. The selection
returned by atomselect is itself a command which you will learn to use.

3 Type set crystal [atomselect top "all"] in the Tk Console window.

This creates a selection containing all the atoms in the molecule and as-
signs it to the variable crystal. Instead of a molecule ID (which is a
number), we have used the shortcut “top” to refer to the top molecule.

The result of atomselect is a function. Thus, $crystal is now a function that

performs actions on the contents of the “all” selection.

4 Type $crystal num. Passing num to an atom selection returns the number
of atoms in that selection. Check that this number matches the number
of atoms for that molecule (as read from the Main form).

5 Type $crystal set beta 0. This resets the “beta” field (which is dis-
played) to be zero for all atoms. As you do this, you should observe that
the atoms on your screen will suddenly change to a uniform color (since
they all have the same beta values).

Atom selections are just references to the atoms in the original molecule. When
you change a property (e.g. beta value) of some atoms through a selection, that
change is reflected in all the other selections that contain those atoms.
2 MULTIPLE MOLECULES AND SCRIPTING 20

6 Now, type set sel [atomselect top "hydrophobic"]. This creates a

selection containing all the hydrophobic residues.

7 Let’s label all hydrophobic atoms by setting their beta values to 1. You
should know how to do this now: $sel set beta 1. If the colors in the
OpenGL Display do not get updated, click on the Apply button at the
bottom of the Graphical Representations window.

Examples of atomic properties. You can get and set many atomic
properties using atom selections, including segment, residue and
atom names, position (x, y and z), charge, mass, occupancy and
radius, just to name a few.

8 You will now change a physical property of the atoms to further illustrate
the distribution of hydrophobic residues. Type $crystal set radius
1.0 to make all the atoms smaller and easier to see through, and then $sel
set radius 1.5 to make the hydrophobic residues larger. The radius field
affects the way that some representations (e.g., VDW, CPK) are drawn.

You have now created a visual state that clearly distinguishes which parts of
the protein are hydrophobic and which are hydrophilic. If you have followed the
instructions correctly, your protein should resemble Fig. 12.

Figure 12: Ubiquitin in the VDW representation, colored according to the hy-
drophobicity of its residues.
2 MULTIPLE MOLECULES AND SCRIPTING 21

Hydrophobic residues. As you probably noticed in your rendering of

ubiquitin, the hydrophobic residues are almost exclusively contained
in the inner core of the protein. This is typical for small water-
soluble proteins. As the protein folds, the hydrophilic residues will
have a tendency to stay at the water interface, while the hydrophobic
residues are pushed together and play a structural role. This helps
the protein achieve proper folding and increases it’s stability.

Atom selections are useful not only for setting atomic data, but also for get-
ting information. Let’s say that you wish to communicate which residues are
hydrophobic, all you need to do is to create a hydrophobic selection and use get
instead of set.

9 Use get with your hydrophobic sel atom selection:

$sel get resname
But there is a problem! Each residue contains many atoms, resulting in multi-
ple repeated entries. Can you think of a way to circumvent this? We know that
each amino acid residue have the same backbone atoms. If you pick only one of
these atoms per residue, each residue will be present only once in your selection.

10 Let’s try this solution. Each residue has one and only one α-carbon
(name CA = alpha):
set sel [atomselect top "hydrophobic and alpha"]
$sel get resname
Aha! It worked.

11 You can also get multiple properties simultaneously. Try the following:
$sel get resid
$sel get {resname resid}
$sel get {x y z}
12 Once you are done with a selection, it is always convinient to delete them
in order to save memory by typing:
$sel delete

2.5 Aligning Two Molecules

To properly visualize how ubiquitin has evolved during the simulation, you
must first make sure to spatially superimpose your two molecules as best as
possible. VMD offers some convenient commands to do this. You will now color
the two molecules differently to visually distinguish them.
2 MULTIPLE MOLECULES AND SCRIPTING 22

1 Make sure that both molecules are “Displayed” (by double-clicking on D

in the VMD Main window).
2 In the Graphical Representations window, change the representation for
the crystal molecule (remember to select it from the menu at the top of
the window). Set the Coloring Method to ColorID, the color to 0 blue and
the Drawing Method to Tube.
3 You can apply changes to different molecules in the Graphical Representa-
tions window by selecting the target molecule from the Selected Molecule
menu at the very top of the window. Choose the same representation for
the molecule called simulation (ColorID and Tube), but set its color to
1 red instead.
4 Create two atom selections containing the protein backbone α-carbons
(using the alpha atom selection macro) for each molecule by typing in the
Tk Console window:
set atomsel1 [atomselect 0 "alpha"]
set atomsel2 [atomselect 1 "alpha"]
Here we have selected the α-carbons because they are more stable than the
protein’s floppy side-chains and give a good indication of the protein’s spatial
conformation.

Even though the two molecules are very different (the crystal structure has no
hydrogens), the selections that you created both contain the exact same atoms.
VMD provides a command – measure fit – for finding the best fit between
two selections corresponding to the same atoms:

measure fit atomsel1 atomsel2 – calculates the best fit matrix

Note atomsel1 and atomsel2 must have the exact same number of atoms. Use
the $atomsel1 num command to check the number.

5 Find the transformation matrix M that will best map the first selection
onto the second by typing:
set M [measure fit $atomsel1 $atomsel2]
6 Apply the matrix you just found to the entire initial molecule (you had
previously defined an atom selection called crystal for this), by typing:
$crystal move $M
In the OpenGL window, the two molecules should now be aligned based on
the positions of their α-carbons. Notice that some parts of the molecule are
well-aligned, whereas other dangling ends and loops seem more floppy (they
have moved more during the equilibration). Realigning the molecules using the
β strands, in addition to the α-helices, results in an alignment resembling Fig 13.
2 MULTIPLE MOLECULES AND SCRIPTING 23

Figure 13: Initial and final states of ubiquitin after spatial alignment.

2.6 Using Color to Display the Deviation

Once the two molecules are aligned, you can compare their atoms’ positions. To
do this, you will use a pre-written script (since it is slightly complex). Feel free
to have a look at the script if you feel adventurous; as you become experienced,
writing such scripts will become trivial. To run a script from a file, you can use
Tcl’s source command:

source file – runs a script from a text file

1 The script is in the coloring.tcl file located in vmd-tutorial-files.

Use the cd command in the VMD TKCon window to get to this directory.
It will change the crystal molecule’s beta values to reflect the displace-
ment (in Å) of each atom of the molecule after equilibration. Let’s run
the script by typing:
source coloring.tcl

2 To see the new beta values, you need to set the Coloring Method of the
crystal molecule to Beta, and hide the simulation molecule (by double-
clicking on D in the VMD Main window).

The crystal molecule should now be colored according to the total displace-
ment that occurred during the simulation. To have a better idea of what went
on, we would like to adjust the color scale in a useful way.

3 In the Graphical Representations window, choose the Trajectory tab. Un-

der the Color Scale Range label, you can set the minimum and maximum
2 MULTIPLE MOLECULES AND SCRIPTING 24

values used for the beta values color scale. Make sure that the right rep-
resentation (of the crystal molecule) is selected, then enter 0 and 5 and
click on Set. This sets the color range to scale between 0 and 5 Å.

Figure 14: The Color Range controls in the Representations window.

4 Now, bring up the Color Controls window (Fig. 15) by choosing the
Graphics → Colors. . . menu item from the VMD Main window. Then
select the Color Scale tab.

Figure 15: The Color Controls window showing the Color Scale tab.

5 In the color scale Method menu, select the BWR color scheme. This will
show residues with a low displacement (0 Å) in blue, those with a high
displacement (5 Å) in red, and those in between as white.

6 You can now also adjust the color scale Midpoint to shift the level of atomic
displacement that will be assigned to white. Try assigning a midpoint of
0.1.

You have now seen how to manipulate continuous color scales on custom data.
Look at your molecule (it should resemble Fig. 16). You should now be able
to determine which parts of ubiquitin are stable (blue) and which are floppy
2 MULTIPLE MOLECULES AND SCRIPTING 25

(red). In general, the dangling ends and loops are flexible, whereas the helices
and β strands are much more rigid.

Figure 16: Ubiquitin colored according to displacement after a 1 ns equilibration.

3 TRAJECTORIES, MACROS AND LABELS 26

3 Trajectories, Macros and Labels

In unit 3, you will learn how to load trajectories, create macros, place labels
on atoms and bonds, and calculate the RMSD of a trajectory using a simple tcl
scripts. At the end, you should determine if the ubiquitin system is equilibrated
by looking at RMSD plots.

3.1 Loading trajectories

You will now learn to load the time evolving coordinates of a system, called
trajectories. You will be able to see a movie of your system.

Trajectory files are normally binary files that contain several sets of coordinates
for the system. Each set of coordinates corresponds to one frame in time. An
example of a trajectory file is a DCD file. The trajectory files do not contain
information of the system contained in the protein structure files (PSF). So we
first need to load the parameter file, and then add the trajectory data to this
file, as explained in Unit 2.

1 Start a new VMD session.

2 Load the PSF file of the system ubiquitin.psf, as done in Unit 2.

3 In the Molecule File Browser window, click on the Browse button. Make
sure that ubiquitin.psf is selected on the menu. Browse for pulling.dcd
in vmd-tutorial-files, click OK, and click on the Load button again.
You will be able to see the frames as they are loaded into the molecule.

4 After the trajectory finishes loading, you will be will be looking at the last
frame of your trajectory. To go to the beginning of the trajectory, you will
use one of the Animation Tools that will be explained extensively later in
the tutorial. In the Main menu, click on the button in the lower left.

5 Choose Graphics → Representations menu item. In Drawing Method, select

NewCartoon, and in the Selected Atoms window, type protein.

6 In the same menu item, create another representation by clicking on the

Create Rep button. In Drawing Method, choose Lines, and in the Selected
Atoms window, type water. For now, turn off this representation by
double-clicking on it.
3 TRAJECTORIES, MACROS AND LABELS 27

Elastic properties of ubiquitin. Ubiquitin has many functions

in the cell. It is currently believed that some of these functions
depend on the elastic properties of ubiquitin.

Only certain proteins are known to show elastic properties.

Examples of these molecules are titin, which acts like a “spring” in
the muscle. The elastic properties of these molecules are due to
hydrogen bonding between residues in β strands of this molecules,
like the ones in ubiquitin.

The trajectory you just loaded is a simulation of an AFM

(Atomic Force Microscopy) experiment pulling on a single ubiquitin
molecule. We will look at the behavior of the protein as it unfolds
while being pulled from one end.

In Molecular Dynamics, before starting a simulation like the

one shown for the AFM experiment, one first needs to perform the
equilibration of the protein. You will have a chance to look at such
an equilibration trajectory later in this tutorial.

In the next section, you will learn how to create representations with a useful
feature called macros. Once you create some representations relevant to this
trajectory, the following section will teach you how to use the Animation Tools
to look at trajectories.

3.2 Macros
You will now create similar representations to the ones you generated in Unit
1. While creating these representations, you will learn macros. A macro is text
that represents a selection. It is useful to create macros when you use certain
selections often. Macros are created with the atomselect command you learned
in unit 2.

atomselect macro name selection – creates a macro for selection

Ubiquitin has a mixed β sheet with five strands. The β sheet play a major role
in the unfolding of the protein. To create a macro for these strands:

1 Open the Tk Console window by choosing the Main → Extensions → Tk

Console.

2 In the Tk Console window, type:

atomselect macro bstrand1 {protein and resid 2 to 6 }

This will create a macro for the first β strand, which includes residues 2 to 6.
3 TRAJECTORIES, MACROS AND LABELS 28

For the other strands, you will find out which residues belong to them by using
the sequence viewer introduced in Unit 1, and then create similar macros:

3 Make sure you are in the first frame of the trajectory, as STRIDE (the
program that calculates secondary structure in VMD) will determine the
structure on that frame.

4 Choose the Extensions → Analysis → Sequence Viewer menu item.

As you learned in Unit 1, the second color column corresponds to structural

features of the protein. The sections in yellow correspond to the β strands.

5 With the mouse, click and drag to highlight the second β strand. This
action will create a representation in the Graphical Representations win-
dow.

6 In the Graphical Representations window, click on the new representation.

The text corresponding to the selection is displayed on the Selected Atoms
window. You should have ( chain U and resid 12 13 14 15 16 ).

7 Use this text to create a new macro by typing in the Tk Console:

atomselect macro bstrand2 { chain U and resid 12 to 16 }

This will create a macro called bstrand2 that will contain residues 12 to 16
of chain U, which corresponds to the protein.

8 Note that the sequence viewer extension locates five β strands. You have
created macros for the first two; create similar macros for the other three
β strands using the sequence extension as done for bstrand2.

Once a macro is created, you can refer to it both in the Tk Console, and in the
Representations selections.

Macros you have created and other macros that come with VMD can be seen
in the Selections tab of the Graphical Representations window. The macros are
listed in the Singlewords window. Clicking on a macro will show its definition
in the Macro Definition window. Double-clicking on it will select it and put its
definition on the Selected Atoms form.

9 Delete the representation created by the sequence viewer.

You will now create a representation with the third and fifth β strands:
3 TRAJECTORIES, MACROS AND LABELS 29

10 In the Graphics → Representations

menu item, click on the Create Rep
button.

11 In the Selected Atoms window,

erase the text that appears there.

12 Click in the Selections tab.

Browse in the Singlewords form
until you find your newly created
macros.

13 Double-click on bstrand3, click

the button or and then double-
click on bstrand5 (Fig. 17).
Then, click on the Apply button.

14 In the Draw Style tab, choose

a NewCartoon representation for
this selection and color it yellow.
You should now see the β sheet.

15 Now, create a similar repre-

sentation with the other three
β strands. Do this by click-
ing on the Create Rep button.
Now, in the Selected Atoms form,
type: bstrand1 or bstrand2 or
bstrand4. Typing them directly
Figure 17: Macros are listed in the Se-
works too!
lections tab of the Graphical Represen-
tations Menu.
3 TRAJECTORIES, MACROS AND LABELS 30

As you can see, macros can be very useful. When saving your work in a saved
state, macros are included in the saved state file.

Macros. You can create several macros and store them in a VMD
preferences file .vmdrc in your home directory (Windows uses the file
vmd.rc). VMD will look for this file upon startup and will recognize
all your macros. For more information of the VMD startup files, refer
to the VMD user’s guide.

To finish this section, you will create a very interesting representation, that
shows a key feature of the trajectory we are looking at. This is an H-bonds
representation.

Hydrogen Bonds. The H-bonds give stability to proteins in several

ways. The H-bonds between β sheet backbone atoms are a trade-
mark of elastic proteins; they give these proteins stability and their
formation and rupture under mechanical stress confers them their
elastic properties.

16 Create a representation with the selection betasheet and backbone, choose

Drawing Method → Hbonds, and color it red selecting Coloring Method →
Color ID. In the options, set Distance Cutoff to 3.2, Angle Cutoff to 30
and Line Thickness to 5.
You can now appreciate the most important features of ubiquitin for this un-
folding trajectory. Your protein should now look similar to the one in Fig. 18.

17 Save a VMD state of this session, so if you want to come back and keep
on working in this tutorial, you don’t have to work through those repre-
sentations again. Do this with the File → Save State... menu item, as
done in Unit 1.
Reusing saved states. When you have a saved state for a system,
you can use it later for viewing different coordinates (PDB or a
different trajectory), if these have the same PSF file. You can do
this by following these steps:

• Load your saved state file: File → Load State...

• Select the file you saved before (i.e. nice-ubiquitin.vmd)
• Go to Molecule → Delete Frames.... You need to delete
all frames currently loaded.The default options will do that.
Click on the Delete button.
• Load your new file yournewtrajectory.dcd into the PSF in
File → Load Data Into Molecule.
3 TRAJECTORIES, MACROS AND LABELS 31

Figure 18: Ubiquitin with key secondary structure features highlighted

3.3 Main Menu Animation Tools

Now that you have nice representations, you will be able to observe features
of your trajectory. The Animation Tools help you do that. The Main Menu
includes all the Animation Tools you need for navigating through your trajec-
tories. They are located at the bottom of the Menu (Fig. 19).

1 Try using the button to jump to the end of the trajectory and go back
to the beginning with the button. You can see the final and initial
states of the trajectory, that correspond to the unfolded and folded states
of the protein.

Figure 19: Animation Tools

2 Turn on again the water representation. Go to the VMD Main window

and choose Graphics → Representations menu item. Double-clicking on the
representation with the text water will turn it on.
3 TRAJECTORIES, MACROS AND LABELS 32

You can click on the slider and drag it back and forth to navigate
through your trajectory. You can stop at anytime you want, or go at the speed
you need. This is helpful when you are looking at a trajectory and want to spot
the time when something interesting happens.

3 Using the slider, observe the behavior of the water around the protein at
the beginning of the trajectory.

Minimizing free energy. Note how the shape of the water box
changes from a cube to a sphere. To minimize free energy, the water
adopts the configuration that exposes less surface to the vacuo, that
is, a sphere! Check how fast the water comes to this configuration
(Each frame step corresponds to 10 ps. )

4 Now, remove the Water Representation from the Graphical Representa-

tions window by double clicking on it to be able to give the protein a closer
look. Slide through the trajectory to look at the protein unfold. Do you
notice any features?

Breaking H-Bonds. Look at the two yellow β strands in the middle.

They are connected to each other with H-bonds. Note how they are
the first β strands to separate, this means their H-bonds are the
first to break. We will talk some more about the relevance of this
phenomenon later.

On the lower part of the Animation tools, you will find all the tools necessary to
play an animation without using the slider. This is done with the Play buttons,
that go forward and backward.

5 Play the trajectory backward. Do you think this is the way the protein
would fold in nature?

There are two ways to change the speed of your animation. You can adjust the
speed of the play using the Speed Slider. You can also adjust the step size. This
is done using the Step Window. If this step is set to 3, the animation will show
every 3rd frame, so it will make it faster.

6 Set the step to 5, and play the trajectory. Note that it plays faster, but it
also looks less smooth than before. However, this can come handy if you
are looking at long trajectories.
3 TRAJECTORIES, MACROS AND LABELS 33

Looping styles. When playing animations, you can choose between

3 looping styles in the Style Chooser. These are “Once”, “Loop”
and “Rock”. A nice trick:

• Go to the beginning of the trajectory (frame 0).

• Set the step number to the total number of frames in your
trajectory (100).
• Set the style to “Rock”.
• Set the speed to lowest.
• Play.
You can see now a comparison between the first and the last frame
of your trajectory. This trick is better than clicking on the Start and
End buttons.

You can also jump to a frame in your trajectory, by entering the frame number
in the window at the left of the Animation Tools.

Recalculate secondary structure. You can ask VMD to recalculate

the secondary structure for a new frame using STRIDE. This was
done for Fig. 18, by typing the command mol ssrecalc top in the
Tk Console, while in frame 28.

NOTE: The Animation Tools you learned cycle through the frames of the Top
molecule, but apply to all Active molecules.

3.4 Labels
In VMD, you can place labels to get information on a particular selection. We
will now make use of those labels for fun and profit. Labels are selected with
the mouse. In this example, we will cover labels that can be placed on atoms
and bonds, although angle and dihedral labeling are also possible.

1 Choose the Mouse → Labels → Atoms menu item. The mouse is now set
to “Display Label for Atom” mode. You can now click on any atom on
your molecule and a label will be placed into this atom. Clicking again on
it will erase the label.

We will now try the same for bonds.

2 Choose the Mouse → Label → Bonds menu item. This selects the “Display
Label for Bond” mode.
3 TRAJECTORIES, MACROS AND LABELS 34

You will make a VDW Representation for the α carbon of Lysine 48 and of the
C terminus. In the pulling simulation, the former is kept fixed, and the latter
is pulled at constant force of 500 pN.

Residue K48. As you learned in Unit 1, polyubiquitin chains can

be linked by a connection between the C terminus of one ubiquitin
molecule and residue K48 of the next. The simulation then mimic
the effect of pulling on the C terminus with this kind of linkage.

To find out the index of these atoms:

3 Make a selection including these two atoms, by typing in the Tk Console:

set sel [atomselect top "resid 48 76 and name CA"]

4 Get the indices:

$sel get index

This command should give the indices 770 1242.

PDB and VMD atom numbering. Note that the atom number of
these atoms in the pdb file is 771 and 1243. VMD starts counting
the index from zero, so the text in the Representation should be
the numbers that VMD understands. This is only the case for index,
since VMD does not read them from the PDB file. Other keywords,
such as residue, are consistent with the PDB file.

5 Create a VDW Representation with selection index 770 1242.

6 Now that you can see them, click on both atoms (one after the other). You
should get a line connecting the two atoms. The number appearing next
to the line is the distance between the two atoms in angstroms (Fig. 20).

The value of the distance corresponds to the current frame.

Labels. The shortcut keys for labels are 1: Atoms and 2: Bonds.
You can use these instead of the Mouse menu. Be sure the Open
GL display window is active when using these shortcuts.
3 TRAJECTORIES, MACROS AND LABELS 35

Figure 20: Bond selection of fixed and pulled atom in simulation. Both atoms
selected display labels in black. The bond is shown in blue, with the value of
the distance between the atoms in angstroms displayed.

Figure 21: Label window.

7 There are more things you can do with labels, in the Graphics → Labels
menu item. In the left side of the window (Fig. 21), there is a pull-down
menu where you can choose the type of label (Atoms, Bonds, Angles,
3 TRAJECTORIES, MACROS AND LABELS 36

Dihedrals). For now, keep it in Atoms. You can see the list of atoms for
which you made a label.

8 Click on one of the atoms. You can see all the information of the atom
displayed. You can delete, hide, or show the label by clicking on these
buttons.

Note that this information is useful to make selections. The information about
the atom corresponds to the current frame, and is updated as the frame is
changed.

9 Now, in the Label window, choose the label type Bonds, and select the
bond you labeled. Note that the information given corresponds to only
the first atom in the bond, but the number in the Value field corresponds
to the length of the bond in angstroms. Click on the Graph tab. Select
the bond you labeled between atoms 770 and 1242. Click on the Graph
button. This will create the plot of the distance between these two atoms
over time. You can also save this data to a file by clicking on the Save
button.

xmgrace. VMD uses the 2-D plotting program xmgrace. This pro-
gram is available from http://plasma-gate.weizmann.ac.il/Grace/.
In Mac OSX, xmgrace runs in the X11 environment. Make sure
X11 is running in order for xmgrace to start.

10 You can now close xmgrace.

Ubiquitin unfolding. The plot displayed shows the length of the

bond over time. You can think of it as a plot of the “length” of the
molecule, as it is pulled at constant force.
Note that at the beginning of the curve, the curve is flattening,
like if the pulling did not affect the structure of the molecule, and
suddenly, the distance increases a lot. Take a look at the time this
occurs: when the two yellow β strands separated! This means that,
those H-bonds that keep the β strands together provide an initial
resistance to the unfolding of the protein, and once they are broken,
the protein unfolds in a more regular way. Try identifying what
happens in the second jump. You will have identified key features
of ubiquitin unfolding!

3.5 An Example Tcl Script: Calculating the RMSD of a

trajectory
VMD is a powerful tool for MD analysis. In this section you will use tcl
scripts to perform analysis of trajectories. You will load a new trajectory, the
3 TRAJECTORIES, MACROS AND LABELS 37

Figure 22: Label window Plot of selected bond over time created with the Graph
button.

equilibration of the ubiquitin system. You will use a short script to calculate
the RMSD of the protein during the MD run, and determine if the system is
equilibrated and ready to simulate.
3 TRAJECTORIES, MACROS AND LABELS 38

Root Mean Square Deviation. The Root Mean Squared Deviation

(RMSD) is a numerical measure of the difference between two
structures. It is defined as :
s
PNatoms
i=1
(ri (t1 ) − ri (t2 ))2
RM SD = (1)
Natoms

where Natoms is the number of atoms whose positions are being

compared, and ri (t) is the position of atom i at time t (or, if you
are comparing two molecules, like in Unit 2, t labels the molecules).

1 Delete the current trajectory. Select the only molecule by clicking it on

the Main window. Go to Molecule → Delete Frames. . . . The default
values on this window will delete the whole trajectory. Click on the Delete
button.

2 Load the equilibration trajectory into the psf file by going to the File → Load
Data Into Molecule. The file is equilibration.dcd.

3 Turn on the water representation and take a look at the trajectory with
the Animation tools that you learned before.

Equilibration. The trajectory you loaded is the equilibration of the

ubiquitin and water system. In reality, this trajectory was simulated
before the pulling trajectory you saw before. In MD, a system
must be equilibrated first to have the stability necessary to observe
other features related to the system and not to changes in the
environment.

Is there anything you can say about this trajectory? Every-

thing seems to be moving around randomly. Note how the water
comes closer to the protein filling in the gaps, and the size of the
water box decreases to obtain the right water density.

Now you will find out how to determine if the protein is equilibrated. One
important factor to determine this is looking at the RMSD of a protein in a
trajectory.

4 The script we are going to use is called rmsd.tcl. This is the script con-
tent:
3 TRAJECTORIES, MACROS AND LABELS 39

set outfile [open rmsd.dat w]

set nf [molinfo top get numframes]
set frame0 [atomselect top "protein and backbone and noh" frame 0]
set sel [atomselect top "protein and backbone and noh"]
set all [atomselect top all]
# rmsd calculation loop
for { set i 1 } { $i <= $nf } { incr i } {
$sel frame $i
$all frame $i
$all move [measure fit $sel $frame0]
puts $outfile "[measure rmsd $sel $frame0]"
}
close $outfile

5 The script does the following:

• Open file rmsd.dat for writing

set outfile [open rmsd.dat w]

• Get the number of frames in the trajectory and assign this value to
the variable nf
set nf [molinfo top get numframes]

• Select the first frame of the molecule to be the one other frames will
compare to. The selection contains the atoms in the backbone of the
protein, excluding hydrogens:
set frame0 [atomselect top "protein and backbone and noh" frame 0]

• Make the same selection as before for an undetermined frame.

set sel [atomselect top "protein and backbone and noh"]

• Make a selection with all atoms.

set all [atomselect top all]

• Text after # denotes comment.

# rmsd calculation loop

• Loop over all frames in the trajectory:

for { set i 1 } { $i <= $nf } { incr i } {
3 TRAJECTORIES, MACROS AND LABELS 40

• Change the selection $sel to the frame to be compared. (Note that

the content of the selection must be identical in every frame, other-
wise a $sel update command needs to be added)
$sel frame $i

• Change the selection $all to the current frame.

$all frame $i

• Calculate the matrix that will fit both selections. Apply this matrix
to the second selection to align the molecules:
$all move [measure fit $sel $frame0]

• Calculate the RMSD value between these two selections, and write
it to file:
puts $outfile "[measure rmsd $sel $frame0]"

You can use the script for the system to test for equilibration.

6 Type source rmsd.tcl in the Tk Console. This will perform all the
commands in the script. The script will write a file rmsd.dat that will
contain the value of the RMSD of the protein backbone against time.

Outside of VMD, you can use some plotting program to see this data. Exam-
ples of these are gnuplot, xmgrace, excel, Mathematica.

7 Use one of the above programs to plot the file rmsd.dat (For example, in
Unix, you can type xmgrace rmsd.dat in a terminal). Can you see the
RMSD curve flattening? This means your system is equilibrated!

More RMSD. You can try sourcing another script called

rmsd-fullthrottle.tcl Take a look at this script to learn simple
procedures in tcl, as well as calculating the RMSD of each residue
over time and coloring residues according to their RMSD similarly
that in Unit 2.

This ends the VMD tutorial. We hope that you learned a lot with it, and that
you will make a great use of all the capabilities VMD has to offer.