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

Pawan Kumar pawan.chinari at gmail.com
Tue Mar 31 14:17:52 CEST 2009


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 for solvation after the genbox step to
remove extra waters from the hydrophobic part of the bilayer ?

Thanking you,

Yours sincerely,
Pawan

On Tue, Mar 31, 2009 at 5:31 PM, Justin A. Lemkul <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>> 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>>> 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>>>> 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> |
>>
>>                   (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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>>
>>    --    ========================================
>>
>>    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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