[gmx-users] PMF of ligand transport

Justin A. Lemkul jalemkul at vt.edu
Mon May 10 15:20:54 CEST 2010



Aswathy wrote:
> 
> 
> On Mon, May 10, 2010 at 6:29 PM, Justin A. Lemkul <jalemkul at vt.edu 
> <mailto:jalemkul at vt.edu>> wrote:
> 
>     Ok . Now I understood. I have one more doubt , as I mentioned to
>     you, I am using one residue in the extracellular loop as a reference
>     point. Since this is in the loop, do you think it can be good
>     reference point (due to th large fluctuatuion in the loop, will it
>     affect the result)?
> 

A residue with a more predictable position is probably preferable.  If the loop 
moves a lot (or even somewhat), the umbrella potential will attempt to maintain 
the specified reference distance, so you could get spurious forces.

-Justin

>     Aswathy wrote:
> 
>         I am pulling through the channel with respect to a single
>         residue on one "side"(extracellular) of the structure. I have
>         used pull_geometry = distance &  pull_dim =  N N Y. From this
>         what I understood is ligand will pull along the z direction with
>         respect to the reference group (away from r_57).  (i.e from
>         extracellular to intracellular). Is this correct?
> 
> 
>     I don't think so.  If you are pulling through a channel, using an
>     extracellular residue as a reference, you will be changing the sign
>     of the distance, rendering "pull_geometry = distance" useless.  For
>     example, in order to properly calculate the PMF, you have to pull
>     from the aqueous solvent, into the channel, then back out into the
>     solvent.  At some point, your ligand is outside the channel (such
>     that, for example, the z-coordinate of the ligand is greater than
>     that of r_57, so distance > 0).  Then, as your ligand enters the
>     channel, its z-coordinate is less than that of r_57, so distance <
>     0.  If this is the case, you must use "pull_geometry = position" to
>     get the correct signs, otherwise your umbrella sampling window
>     reference distances will be nonsensical.
> 
>     -Justin
> 
>         Here is my umbrella sampling .mdp parameters
> 
>         pull                     = umbrella
>         pull_geometry            = distance
>         pull_dim                 =  N N Y
>         pull_start               = yes
>         pull_nstxout             =  10
>         pull_nstfout             =  10
>         pull_ngroups             =  1
>         pull_group0              =  r_57
>         pull_group1              =  r_C1
>         pull_k1                  =  1000
>         pull_init1               =  0
> 
>         On Mon, May 10, 2010 at 4:50 PM, Justin A. Lemkul
>         <jalemkul at vt.edu <mailto:jalemkul at vt.edu>
>         <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>>> wrote:
> 
> 
> 
>            Aswathy wrote:
> 
>                Thanks for your  reply.
> 
>                 In this case reference (r57) is not the part of the channel.
>                But it is a residue in the loop above the channel entry.
>         Thats
>                why I used pull_geometry=distance. Therefore I am pulling the
>                ligand away from this reference.
> 
> 
>            So you are not pulling through the channel?  Or you are pulling
>            through the channel with respect to a single residue on one
>         "side"
>            of the structure?  If your ligand ever crosses over this
>         reference
>            in any way, the reference distance will change sign and thus
>         Tom is
>            right, you should use "pull_geometry = position."  With
>         "distance,"
>            you can only ever have positive reference distances.
> 
>            What are your .mdp settings during umbrella sampling?
> 
>            -Justin
> 
>                Thanks
>                -Aswathy
> 
> 
>                On Mon, May 10, 2010 at 3:05 PM, Thomas Piggot
>                <t.piggot at soton.ac.uk <mailto:t.piggot at soton.ac.uk>
>         <mailto:t.piggot at soton.ac.uk <mailto:t.piggot at soton.ac.uk>>
>                <mailto:t.piggot at soton.ac.uk
>         <mailto:t.piggot at soton.ac.uk> <mailto:t.piggot at soton.ac.uk
>         <mailto:t.piggot at soton.ac.uk>>>> wrote:
> 
>                   Hi,
> 
>                   If you defined the reference (r_57) as part of your
>         channel then
>                   with pull_geometry=distance you will have problems as the
>                distance
>                   between pull_group1 and pull_group0 becomes closer to zero
>                and then
>                   the distance becomes positive again.
> 
>                   I recently had this with my umbrella sampling
>         simulations. Search
>                   for the discussion of things you can do to address
>         this issue
>                on the
>                   list. To stop this being a problem in the first place
>         you should
>                   have used pull_geometry=position.
> 
>                   Cheers
> 
>                   Tom
> 
>                   Aswathy wrote:
> 
>                       Can any one help me please? I looking forward to hear
>                from any
>                       of you.
>                       Thank you.
> 
> 
>                       On Thu, May 6, 2010 at 1:19 PM, Aswathy
>                <ammasachu at gmail.com <mailto:ammasachu at gmail.com>
>         <mailto:ammasachu at gmail.com <mailto:ammasachu at gmail.com>>
>                       <mailto:ammasachu at gmail.com
>         <mailto:ammasachu at gmail.com> <mailto:ammasachu at gmail.com
>         <mailto:ammasachu at gmail.com>>>
>                <mailto:ammasachu at gmail.com <mailto:ammasachu at gmail.com>
>         <mailto:ammasachu at gmail.com <mailto:ammasachu at gmail.com>>
> 
>                       <mailto:ammasachu at gmail.com
>         <mailto:ammasachu at gmail.com>
>                <mailto:ammasachu at gmail.com
>         <mailto:ammasachu at gmail.com>>>>> wrote:
> 
>                          Ok i will explain you in detail.
> 
>                           Initially i pulled the ligand through the protein
>                channel ,
>                       using
>                          the given parameters.
> 
>                          pull                     = umbrella
>                          pull_geometry            = distance
>                          pull_dim                 =  N N Y
>                          pull_start               = yes
>                          pull_nstxout             =  10
>                          pull_nstfout             =  10
>                          pull_ngroups             =  1
>                          pull_group0              =  r_57
>                          pull_group1              =  r_C1
>                          pull_rate1               =  0.01
>                          pull_k1                  =  1500
> 
>                          Then I extracted the frames from the trajectory
>         using
>                the perl
>                          program provided with tutorial. COM distance I
>         took as
>                nearly
>                       0.12
>                          nm. (But sometimes I failed to obtain frames
>         exactly
>                at that
>                          interval, but took  nearly at 0.12). Each frame
>         I used for
>                       Umbrella
>                          sampling for 1ns.
>                          Then I checked histograms for overlapping (Some
>                histograms were
>                          entirely overlapped and I removed that from the
>         list,
>                where ever
>                          gaps i selected new frames and did sampling so
>         that I
>                can get an
>                          evenly distributed histograms , I know this will
>                change the
>                       overall
>                          COM distribution but is there any other way to
>         solve
>                this?) .
> 
>                          Finally once I obtained reasonably good overlapped
>                histograms, I
>                          plotted PMF using g_wham. The plot  was a steeply
>                increasing
>                          potential.  How can we get increased PMF even
>         when the
>                ligand is
>                          reached out of the channel?
> 
> 
>                                   Did I made any mistake any where , I
>         am confused.
> 
>                          Thank you.
>                          -Aswathy
> 
> 
> 
>                          On Thu, May 6, 2010 at 12:56 PM, Jochen Hub
>                       <jochen at xray.bmc.uu.se
>         <mailto:jochen at xray.bmc.uu.se> <mailto:jochen at xray.bmc.uu.se
>         <mailto:jochen at xray.bmc.uu.se>>
>                <mailto:jochen at xray.bmc.uu.se
>         <mailto:jochen at xray.bmc.uu.se> <mailto:jochen at xray.bmc.uu.se
>         <mailto:jochen at xray.bmc.uu.se>>>
>                          <mailto:jochen at xray.bmc.uu.se
>         <mailto:jochen at xray.bmc.uu.se>
>                <mailto:jochen at xray.bmc.uu.se <mailto:jochen at xray.bmc.uu.se>>
>                       <mailto:jochen at xray.bmc.uu.se
>         <mailto:jochen at xray.bmc.uu.se>
>                <mailto:jochen at xray.bmc.uu.se
>         <mailto:jochen at xray.bmc.uu.se>>>>> wrote:
> 
>                              Aswathy wrote:
> 
> 
>                                  Hi gromacs users,
> 
>                                  I am using Gromacs 4.0.4 package. I am
>         doing
>                SMD of a
>                       ligand
>                                  transport through a channel.
> 
>                                  I performed SMD and did umbrella sampling
>                (Thanks to
>                       Justin
>                                  for his  tutorial). Extracted frames
>         with a window
>                       spacing
>                                  interval  of ~0.12nm. and did 1ns sampling.
>                       Histograms are
>                                  with reasonabvle overlap. Then I used
>         g_wham
>                for plotting
>                                  PMF considering first 300ps as
>         equilibration.
> 
>                              Isn't SMD usually referred to pulling at some
>                finite pulling
>                              speed? That would not be umbrella sampling.
> 
>                              Anyway, you'll have to provide a lot more
>         data to
>                enable
>                       us to
>                              help you.
> 
>                              Jochen
> 
> 
> 
> 
>                                  I am getting a plot , but potential is
>         increasing
>                                  constantly. ie, PMF is not converged as
>                mentioned the
>                                  tutorial? Do I need to extend the
>         sampling ?
>                or any other
>                                  reason?
> 
>                                  Please help me.
>                                  Thank you.
> 
>                                  -Aswathy
> 
> 
> 
>                              --                      
>         ---------------------------------------------------
>                              Dr. Jochen Hub
>                              Molecular Biophysics group
>                              Dept. of Cell & Molecular Biology
>                              Uppsala University. Box 596, 75124 Uppsala,
>         Sweden.
>                              Phone: +46-18-4714451 Fax: +46-18-511755
>                            
>          ---------------------------------------------------
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> 
> 
>                          --     Aswathy
> 
> 
> 
> 
>                       --         Aswathy
> 
> 
>                   --     Dr Thomas Piggot
>                   University of Southampton, UK.
> 
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> 
> 
>                --         Aswathy
> 
> 
>            --     ========================================
> 
>            Justin A. Lemkul
>            Ph.D. Candidate
>            ICTAS Doctoral Scholar
>            MILES-IGERT Trainee
>            Department of Biochemistry
>            Virginia Tech
>            Blacksburg, VA
>            jalemkul[at]vt.edu <http://vt.edu> <http://vt.edu> | (540)
>         231-9080
> 
>            http://www.bevanlab.biochem.vt.edu/Pages/Personal/justin
> 
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> 
> 
>         -- 
>         Aswathy
> 
> 
>     -- 
>     ========================================
> 
>     Justin A. Lemkul
>     Ph.D. Candidate
>     ICTAS Doctoral Scholar
>     MILES-IGERT Trainee
>     Department of Biochemistry
>     Virginia Tech
>     Blacksburg, VA
>     jalemkul[at]vt.edu <http://vt.edu> | (540) 231-9080
>     http://www.bevanlab.biochem.vt.edu/Pages/Personal/justin
> 
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> 
> -- 
> Aswathy

-- 
========================================

Justin A. Lemkul
Ph.D. Candidate
ICTAS Doctoral Scholar
MILES-IGERT Trainee
Department of Biochemistry
Virginia Tech
Blacksburg, VA
jalemkul[at]vt.edu | (540) 231-9080
http://www.bevanlab.biochem.vt.edu/Pages/Personal/justin

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