[gmx-users] Re: Doubt regarding membrane protein in POPC bilayer

Justin A. Lemkul jalemkul at vt.edu
Tue Mar 31 14:20:37 CEST 2009



Pawan Kumar wrote:
> Respected Sir,
> 
> Greetings from Pawan.
> Thanks for all your kind help and suggestions.
> I will work on this and ask you if I have further doubts.
> Is it fine if I use the perl code given in 
> wiki.gromacs.org/membrane-simulations 
> <http://wiki.gromacs.org/membrane-simulations> for solvation after the 
> genbox step to remove extra waters from the hydrophobic part of the 
> bilayer ?
> 

You mean the C program or shell script at 
http://wiki.gromacs.org/index.php/Membrane_Simulations?  Yes, either of those 
should be fine.

-Justin

> Thanking you,
> 
> Yours sincerely,
> Pawan
> 
> On Tue, Mar 31, 2009 at 5:31 PM, Justin A. Lemkul <jalemkul at vt.edu 
> <mailto:jalemkul at vt.edu>> wrote:
> 
> 
> 
>     Pawan Kumar wrote:
> 
>         Respected Sir,
> 
>         Greetings from Pawan.
>         Thanks for all your suggestions.
>         Is it possible to use the lipid.itp file from Tieleman sir's
>         website in combination with GROMOS 96 force field without any
>         modification ?
> 
> 
>     If you want to use lipid.itp without any modification, you are
>     restricted to the ffgmx (deprecated!) force field.  For
>     Gromos96-lipid.itp combination, simply remove the ";; parameters for
>     lipid-GROMOS interactions," as these nonbonded combinations are for
>     ffgmx only.  Otherwise, the nonbonded interactions (which now have
>     consistent combination rules) should be generated correctly.
> 
> 
>         Is it fine if I use the Gromos/Berger force field combination
>         for the system I am using ?
> 
> 
>     That is a decision you will have to make based on a thorough
>     examination of the literature, and the benefits and criticisms of
>     these particular force field parameters.
> 
> 
>         I am sorry to ask this but can you please help me with some
>         information how to modify the lipid.itp file ?
> 
> 
>     That depends entirely upon what you want to do.  If you want
>     Gromos/Berger, I've already told you.  If you want OPLS/modified
>     Berger, search the archives for Chris Neale's posts on that topic.
> 
>     -Justin
> 
>         I will edit vdwradii.dat file as per your suggestion.
> 
> 
>         Thanking you,
> 
>         Yours sincerely,
>         Pawan
> 
>         On Tue, Mar 31, 2009 at 4:53 PM, Justin A. Lemkul
>         <jalemkul at vt.edu <mailto:jalemkul at vt.edu>
>         <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>>> wrote:
> 
> 
> 
>            Pawan Kumar wrote:
> 
>                Respected Sir,
> 
>                Greetings from Pawan.
>                I have edited the lipid.itp file to add just one line
>         extra " H
>                atom from the opls_force_filed.itp " at the end of lipid
>                interactions data and that works fine.
>                I have done this after seeing the archives. It was given
>         either
>                I should change the file for sigma and epsilon values or else
>                just add this line.
>                I think I have done it correctly. If not please correct me.
> 
> 
>            No, that is incorrect.  You need to add the additional atom
>         type as
>            well as convert C6/C12 to sigma/epsilon.  Otherwise, the
>         calculated
>            nonbonded interactions are meaningless.  Read instructions
>         carefully!
> 
>            Force fields have to be internally self-consistent, so using the
>            parameters from OPLS with Berger lipids will give spurious
>         results.
>             The only proper combinations are Gromos/Berger or OPLS/converted
>            Berger.
> 
> 
>                I have removed the position restraints after the inflategro
>                procedure.
>                After that I did energy minimization using define =
>         -DFLEXIBLE.
>                I will change the temperature as per your suggestion.
>                I thought before solvating if I do position restraint
>         mdrun the
>                lipids will compact more surrounding the protein.
> 
> 
>            No, your system will probably explode due to unsatisfied charge
>            interactions and hydrogen bonds.  There is a substantial
>         dipole in
>            each lipid headgroup that can repel the lipids away from each
>         other
>            if it is not shielded.
> 
> 
>                Thats the step when I got Lincs warnings and segmentation
>         fault.
>                Then I tried solvating the system using genbox step and
>                spc216.gro as the solvent.
>                But before doing the solvation step I copied the vdwradii.dat
>                file into the working directory and increased the value for
>                carbon to 0.5.
>                But the result of this was " Segmentation fault " again.
>         Can you
>                please tell me why I get the " Segmentation fault " here
>         in this
>                step.
>                The command used was : genbox  -cp comp_em27.pdb -cs
>         spc216.gro
>                -o box.pdb -p topol.top
> 
> 
>            Using a 0.5-nm radius for carbon will cause problems of excess
>            memory consumption, or otherwise breaks the calculation.  Use
>            something more along the lines of 0.35 or 0.375, and manually
>         delete
>            out any stray waters in the hydrophobic core.
> 
>            -Justin
> 
> 
>                Thanking you,
> 
>                Yours sincerely,
>                Pawan
> 
>                On Tue, Mar 31, 2009 at 4:20 PM, Justin A. Lemkul
>                <jalemkul at vt.edu <mailto:jalemkul at vt.edu>
>         <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>>
>                <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>
>         <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>>>> wrote:
> 
> 
> 
>                   Justin A. Lemkul wrote:
> 
> 
> 
>                       Pawan Kumar wrote:
> 
>                           Respected Sir,
> 
>                           Greetings from Pawan.
>                           I have used force constants of 100000 in position
>                restraint
>                           .itp files for proteins as suggested in Dr.
>         Tieleman' s
>                           webisite for Inflategro.
>                           The mdp files are :
> 
> 
>                       The .mdp files look reasonable enough, although I
>         don't
>                know why
>                       you are applying position restraints during EM.
>          If it is for
>                       InflateGRO, that is fine, but once the system is
>                assembled, you
>                       should remove the position restraints from the
>         protein to
>                       minimize the system more.
> 
>                       And are you sure you want 300K?  DPPC will be in a gel
>                phase at
>                       that temperature.  If you want a more realistic
>         fluid-phase
>                       model, you'll have to go above 315K (323K is common).
> 
> 
>                   One thing I just noticed.  You don't have solvent in your
>                   position-restrained run?  That could be a big problem
>         if the
>                lipid
>                   headgroups are strongly repelled from one another.
>          Add solvent
>                   before doing anything other than EM.
> 
>                   -Justin
> 
> 
>                           *Topology file :*
>                           ; Include forcefield parameters
>                           #include "ffoplsaa.itp"
>                           #include "lipid.itp"
>                           #include "dppc.itp"
> 
> 
>                       This section should not work, as written.  Have
>         you modified
>                       lipid.itp according to Chris Neale's half-epsilon
>                       double-pairlist method?  If not, what you've done
>         makes no
>                       sense.  The Berger lipid parameters distributed
>         through
>                       Tieleman's site are designed for use with the
>         Gromos force
>                       fields.  They can be modified (search in the
>         archives),
>                but that
>                       can also be a source of error.  Users who have made
>                mistakes in
>                       the conversion have seen their systems explode.
> 
>                       -Justin
> 
>                           ; Include chain topologies
>                           #include "topol_A.itp"
>                           #include "topol_B.itp"
>                           #include "topol_C.itp"
> 
>                           ;#ifdef POSRES
>                           ;#include "lipid_posre.itp"
>                           ;#endif
> 
>                           ; Include water topology
>                           #ifdef FLEX_SPC
>                           #include "flexspc.itp"
>                           #else
>                           #include "spc.itp"
>                           #endif
> 
>                           #ifdef POSRES_WATER
>                           ; Position restraint for each water oxygen
>                           [ position_restraints ]
>                           ;  i funct       fcx        fcy        fcz
>                             1    1       1000       1000       1000
>                           #endif
> 
>                           ; Include generic topology for ions
>                           #include "ions.itp"
> 
>                           [ system ]
>                           ; Name
>                           PROTEIN IN DPPC BILAYER
> 
>                           [ molecules ]
>                           ; Compound      #mols
>                           Protein_A           1
>                           Protein_B           1
>                           Protein_C           1
>                           DPPC              926
>                           ;SOL             23552
> 
>                           Please help with some suggestions.
> 
>                           Thanking you,
> 
>                           Yours sincerely,
>                           Pawan
> 
>                           On Mon, Mar 30, 2009 at 6:22 PM, Justin A. Lemkul
>                           <jalemkul at vt.edu <mailto:jalemkul at vt.edu>
>         <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>>
>                <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>
>         <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>>>
>                           <mailto:jalemkul at vt.edu
>         <mailto:jalemkul at vt.edu> <mailto:jalemkul at vt.edu
>         <mailto:jalemkul at vt.edu>>
>                <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>
>         <mailto:jalemkul at vt.edu <mailto:jalemkul at vt.edu>>>>> wrote:
>                                          Pawan Kumar wrote:
> 
>                                  Respected Sir,
> 
>                                  Greetings from Pawan.
>                                  I did the Inflategro procedure for the POPC
>                bilayer
>                           generated
>                                  using genconf.
>                                  It took around 26 compressions for
>         coming near the
>                           initial area
>                                  (just above it).
>                                  The minimization were all converged to
>         Fmax <
>                1350.
>                                  If I decrease the Fmax less than this I
>         am getting
>                           machine
>                                  precision.
>                                  But when I proceeded with the final
>         compressed
>                           structure for pr
>                                  mdrun it gave lincs warnings and ended with
>                           segmentation fault.
>                                  As an alternative to this bilayer I
>         used the
>                DPPC bilayer
>                                  (pre-equilibrated) which is given in
>         GMX-Benchmark
>                           distribution.
>                                  I carried out the same steps of
>         Inflategro. I used
>                           the cutoff of
>                                  14 A in the inflation and compression
>         step also.
>                                  In this case the the Steepest Descents
>                converged to
>                           Fmax < 1000
>                                  in all the steps. The maximum force war
>         never
>                above 850.
>                                  But when I did position restraint mdrun
>         with
>                the last
>                           compressed
>                                  file I got Lincs warnings and
>         Segementation fault
>                           after few
>                                  steps (30 - 40 steps).
>                                  Can you please help how to proceed ?
> 
> 
>                              If the minimization procedure is adequately
>         finishing,
>                           then the
>                              problem comes from something you are doing.  If
>                you post
>                           your .mdp
>                              file, we may be able to see if there are
>         any obvious
>                           mistakes.
> 
>                              -Justin
> 
>                                  Thanking you,
> 
>                                  Yours sincerely,
>                                  Pawan
> 
> 
>                                  Justin A. Lemkul
>                                  Graduate Research Assistant
>                                  ICTAS Doctoral Scholar
>                                  Department of Biochemistry
>                                  Virginia Tech
>                                  Blacksburg, VA
>                                  jalemkul[at]vt.edu <http://vt.edu>
>         <http://vt.edu>
>                <http://vt.edu> <http://vt.edu> |
> 
>                           (540) 231-9080
>                                      
>          http://www.bevanlab.biochem.vt.edu/Pages/Personal/justin
> 
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> 
> 
>                   --    ========================================
> 
>                   Justin A. Lemkul
>                   Graduate Research Assistant
>                   ICTAS Doctoral Scholar
>                   Department of Biochemistry
>                   Virginia Tech
>                   Blacksburg, VA
>                   jalemkul[at]vt.edu <http://vt.edu> <http://vt.edu>
>         <http://vt.edu> | (540)
>                231-9080
>                   http://www.bevanlab.biochem.vt.edu/Pages/Personal/justin
> 
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>            --    ========================================
> 
>            Justin A. Lemkul
>            Graduate Research Assistant
>            ICTAS Doctoral Scholar
>            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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> 
> 
> 
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>     -- 
>     ========================================
> 
>     Justin A. Lemkul
>     Graduate Research Assistant
>     ICTAS Doctoral Scholar
>     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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-- 
========================================

Justin A. Lemkul
Graduate Research Assistant
ICTAS Doctoral Scholar
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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