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POVME2 (POcket Volume MEasurer 2) scans a molecular-dynamics simulation and extracts druggable protein pockets with unique conformations. It is useful because ligand-binding pockets adopt many conformations in response to atomic forces. Characterizing these pocket geometries helps identify those that are particularly amenable to ligand binding.

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POVME 2.2.2

0. License: GNU General Public License version 3

This program is free software: you can redistribute it and/or modify it under the terms of the GNU General Public License as published by the Free Software Foundation, either version 3 of the License, or (at your option) any later version.

This program is distributed in the hope that it will be useful, but WITHOUT ANY WARRANTY; without even the implied warranty of MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU General Public License for more details.

You should have received a copy of the GNU General Public License along with this program. If not, see https://www.gnu.org/licenses/.

1. Download POVME 2.2.2

Begin by downloading POVME 2.2.2. An example input file is included with the download (in the 'povme/examples/' directory). This input file is heavily commented and may be even more useful that the information provided on this website.

POVME is released under the GNU General Public License. If you have any questions, comments, or suggestions, please don't hesitate to contact me, Jacob Durrant, at durrantj [at] pitt [dot] edu.

If you use POVME in your work, please cite:

  1. Durrant, J. D., C. A. de Oliveira, et al. (2011). "POVME: An algorithm for measuring binding-pocket volumes." J Mol Graph Model 29(5): 773-776.
  2. Durrant, J.D., L. Votapka, J. Sørensen, and R. E. Amaro (2014). "POVME 2.0: An Enhanced Tool for Determining Pocket Shape and Volume Characteristics." J. Chem. Theory Comput. 10(11):5047-5056.

2. Align a PDB-formatted trajectory

POVME accepts a multi-frame PDB (Protein Data Bank) file as input. The computer program Visual Molecular Dynamics (VMD) is useful for aligning trajectories and converting files to the PDB format. Alignment is necessary because the POVME algorithm assumes the pocket being measured does not translate or rotate in space. We note also that single-frame PDB files can likewise serve as POVME input if the user wishes only to measure the volume of a single pocket.

3. Define an inclusion region

The user must define an "inclusion" region. This region is constructed from a combination of user-specified spheres and rectangular prisms. The required inclusion region should entirely encompass all the binding-pocket conformations of the trajectory. Specify the spheres and rectangular prisms of the inclusion region in a text-based POVME input file (e.g., "POVME.in"):

PointsInclusionSphere -7.12 2.60 -4.67 6.0
PointsInclusionSphere -2.0 -2.0 -4.0 5.0
PointsInclusionBox -5.0 -7.0 2.0 10.0 10.0 10.0
PointsInclusionBox -2.0 -9.0 -3.0 10.0 5.0 10.0

Note that for the spheres, the first three parameters are coordinates, and the fourth parameter is a radius. For the rectangular prisms (i.e. "boxes"), the first three parameters are coordinates, and the last three are the dimensions of the box in the X, Y, and Z directions.

4. Define an exclusion region

An optional exclusion region defines portions of the inclusion region that should be ignored, perhaps because they are not truly associated with the pocket. It is similarly constructed from spheres and boxes:

PointsExclusionSphere -2.0 -2.0 -4.0 5.0
PointsExclusionBox -5.0 -7.0 2.0 10.0 10.0 10.0

5. Create a field of equidistant points

To generate a field of equidistant points that encompasses all the binding-pocket conformations of the trajectory, POVME first floods the user-specified inclusion region with points and then removes any points also contained in the optional exclusion region. You need to specify the distance separating each of these equidistant points:

GridSpacing 1.0

6. How to choose the inclusion and exclusion regions

As you can imagine, identifying just the right set of inclusion and exclusion spheres and boxes to encompass the binding pocket is challenging. One approach is to define an initial geometry, visualize that geometry together with the receptor using a program like VMD, and then iteratively add new inclusion and exclusion regions as required. You can optionally save the point field to a file called point_field.pdb for visualization:

SavePoints true

7. Specify the location of the receptor PDB file to analyze

Once you've properly generated a pocket-encompassing point field, you're ready to use that point field to calculate pocket volumes. Here's how to specify the location of the PDB receptor file that has the pocket you wish to analyze:

PDBFileName my_receptor.pdb

Note that this file can be a trajectory containing multiple frames.

8. Remove points that are near receptor atoms

As the purpose of POVME is to measure the volume of a binding-pocket cavity, the program next removes any points that are close to receptor atoms, leaving only those points that are likely to be located within the binding pocket itself. To specify how close the points can come to the van der Waal's surface of the receptor before being removed:

DistanceCutoff 1.09

Note that if the receptor PDB file contains multiple frames, this will be done on a frame-by-frame basis.

9. Remove points outside the receptor's convex hull

POVME 2.0 introduced an optional new feature for removing points that lie entirely outside the binding pocket. Specifically, the gift-wrapping algorithm is used in combination with the Akl-Toussaint heuristic to define the convex hull of receptor atoms near the user-defined inclusion region. Any points that fall outside the convex hull are removed. This feature is particularly useful when the user defines an inclusion region that protrudes into the surrounding solvent-occupying space. To activate the convex-hull feature:

ConvexHullExclusion true

10. Remove points that are not contiguous with the primary pocket

Like the original POVME program, version 2.2.2 retains the optional ability to remove isolated patches of points that are not contiguous with the primary binding pocket. POVME perfoms this task in two steps.

First, POVME applys a low-pass filter to check the connectedness of the remaining pocket-occupying points. It retains only those points that have at least X neighboring points, where X is defined:

ContiguousPointsCriteria 3

(Note that points that are "kitty-corner" from each other count as neighbors.)

Second, POVME identifies which of the remaining connected points are also connected to a user-defined contiguous pocket seed volume (e.g., located at the center of the traditionally defined pocket), where the seed volume is defined:

ContiguousPocketSeedSphere 67.0 102.0 57.0 4.0
ContiguousPocketSeedBox 50.0 50.0 50.0 10.0 10.0 10.0

All pocket-occupying points contiguous (or connected) to this region are retained, but isolated patches of points that are not directly connected are deleted.

11. Additional POVME parameters

Here are some additional POVME parameters you might find helpful:

# Tell POVME how to perform the calculations.

NumProcessors               12                             # POVME can use multiple processors on
                                                           # Unix-based systems.

UseDiskNotMemory            false                          # In some cases, your PDB trajectory may
                                                           # be so large that the resulting POVME
                                                           # analysis cannot be easily stored in
                                                           # your computer's memory. If
                                                           # UseDiskNotMemory is set to true, POVME
                                                           # will rely more on your disk space than
                                                           # on memory/RAM.

# Tell POVME how to save the output

OutputFilenamePrefix          ./POVME_test_run/POVME_      # All the files POVME outputs will start
                                                           # with this prefix. POVME automatically
                                                           # creates any required directory
                                                           # (./POVME_test_run/ in this case).

SaveIndividualPocketVolumes   true                         # If true, POVME saves the pocket-volume
                                                           # points of each frame to a separate PDB
                                                           # file. The file names will be like
                                                           # {PREFIX}frame_X.pdb.

SavePocketVolumesTrajectory   true                         # If true, POVME saves all the pocket-
                                                           # volume points of each frame to a single
                                                           # PDB trajectory file. The individual
                                                           # frames are separated by END cards. The
                                                           # file name will be
                                                           # {PREFIX}volume_trajectory.pdb.

OutputEqualNumPointsPerFrame  true                         # Some visualization programs (e.g. VMD)
                                                           # are only compatible with trajectories
                                                           # that have the same number of atoms in
                                                           # each frame. If EqualNumAtomsPerFrame is
                                                           # true, POVME adds extra points at the
                                                           # origin (0.0, 0.0, 0.0) to satisfy this
                                                           # requirement. This affects files created
                                                           # with both SaveIndividualPocketVolumes
                                                           # and SavePocketVolumesTrajectory but
                                                           # does not alter the volume calculation
                                                           # itself.

SaveTabbedVolumeFile          true                         # If true, POVME saves the calculated
                                                           # volumes to a file in a simple tabular
                                                           # format that can be easily pasted into
                                                           # popular spreadsheet programs like
                                                           # Microsoft Excel. The file is named
                                                           # {PREFIX}volumes.tabbed.txt

SaveVolumetricDensityMap      true                         # If true, POVME saves a volumetric
                                                           # density map in the DX format. A
                                                           # volumetric density value is associated
                                                           # with each of the pocket-occupying
                                                           # points by calculating the fraction of
                                                           # all trajectory pocket volumes that
                                                           # include the given point. The file is
                                                           # named {PREFIX}volumetric_density.dx.

CompressOutput                true                         # If you're short on disk space, POVME
                                                           # can automatically compress all output
                                                           # files using gz compression.

12. POVME output

By default, POVME writes a number of files to the disk. The calculated pocket volumes, as well as user-defined parameters and progress messages, are saved to a simple text-based log file. POVME can also be instructed to save the volume measurements to a second file in a simple tabular format that can be easily pasted into popular spreadsheet programs. Pocket-occupying points are equidistant (1.0 Å by default), so each point is associated with an identical cubical volume (e.g. 1.0 Å3). The volume of a whole pocket is calculated by simply summing the individual volumes associated with each unique point.

POVME also optionally saves the pocket-occupying points of each frame to PDB file(s) on this disk. The user can instruct the program to save these points to separate files and/or to a single PDB trajectory. Some visualization programs (e.g. VMD) are only compatible with trajectories that have the same number of atoms in each frame. POVME can optionally write extra points to the origin (0.0, 0.0, 0.0) on a frame-by-frame basis to satisfy this requirement.

Finally, POVME also optionally saves a volumetric density map in the Data Explorer (DX) format. A volumetric density value is associated with each of the pocket-occupying points by calculating the fraction of all trajectory pocket volumes that include the given point. If the density map is displayed as an isosurface, the value of the isosurface expresses the fraction of time (e.g. over the course of the simulation) that the pocket included the displayed volume.

(README.md adapted from this website.)

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POVME2 (POcket Volume MEasurer 2) scans a molecular-dynamics simulation and extracts druggable protein pockets with unique conformations. It is useful because ligand-binding pockets adopt many conformations in response to atomic forces. Characterizing these pocket geometries helps identify those that are particularly amenable to ligand binding.

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