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

Justin A. Lemkul jalemkul at vt.edu
Mon Mar 23 13:06:10 CET 2009



Pawan Kumar wrote:
> Respected Sir,
> 
> Greetings from Pawan.
> Thanks for your reply.
> I used to do genbox first and then Inflategro all the time.
> Is it possible to concatenate one file after the other using " vi 
> editor" - I mean just copying the contents of the membrane.gro file at 
> the end of protein.gro file ?

cat protein.gro membrane.gro > system.gro

> Is it fine if I generate the .gro files using editconf command ?

Yes, you will have to center the protein and membrane appropriately, within a 
box that has the same dimensions.

-Justin

> Thanks for your kind help.
> 
> Thanking you,
> Pawan
> 
> On Mon, Mar 23, 2009 at 5:02 PM, Justin A. Lemkul <jalemkul at vt.edu 
> <mailto:jalemkul at vt.edu>> wrote:
> 
> 
> 
>     Pawan Kumar wrote:
> 
>         Respected Sir,
> 
>         Greetings from Pawan.
>         Thanks for your mail.
>         I will tell you in detail.
>         The control lipid bilayer ( which is generated using genconf
>         -nbox 2 2 1 -dist 0 0 0 ) minimized fine with an emtol value of
>         1510.
>         Then after inserting the protein with genbox the whole system
>         minimized with an emtol value of 2250.
>         Then I am trying to do position restraint mdrun where I am
>         getting errors.
>         Inflategro can be used only after inserting the protein right
>         (which I did using genbox with -vdwd option as 0.6) ?
> 
> 
>     No.  Read the documentation carefully.  You first concatenate your
>     protein and membrane .gro files, then use InflateGRO on that system.
>      There is no genbox involved when using InflateGRO, hence why I
>     suggested it as a (potentially) more reliable method.
> 
>     -Justin
> 
>         Thanking you,
>         Pawan
> 
> 
>         On Mon, Mar 23, 2009 at 4:39 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:
> 
>                Hello Justin Sir,
> 
>                Greetings from Pawan.
>                Sorry for the inconvenience.
>                Next time I will keep in mind about the subject line.
>                I tried deleting those particular atoms where it gave the
>                maximum force.
> 
> 
>            You said that this atom was part of a lipid.  Deleting these
>         atoms
>            will give you a broken structure and an unrealistic system.  It
>            should also cause grompp to complain.
> 
> 
>                But every time a new atom with a maximum force was given
>         in the
>                output.
>                Is it fine to use to use "constraints = all-angles" in
>         order to
>                overcome the Lincs warnings ?
> 
> 
>            No.  LINCS warnings indicate that there is something
>         fundamentally
>            wrong with your system.  It likely stems from how you
>         inserted your
>            protein into the lipids.  Think about it.  Your control lipid
>            bilayer minimized fine.  Your protein-membrane system does
>         not, if
>            that is what we're still talking about. That indicates to me that
>            the problem is not with the lipids, the problem is with the
>            insertion of the protein.
> 
>            Instead of genbox (which is somewhat crude), try the InflateGRO
>            script from Tieleman's site.  I have used it many times to build
>            nicely-packed membrane protein systems.  It takes substantially
>            longer to build the system than with genbox, but I think the
>         results
>            are more reliable.
> 
>            -Justin
> 
>                Thanks in advance.
> 
>                Thanking you,
>                Pawan
> 
> 
> 
> 
>                On Mon, Mar 23, 2009 at 4:16 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:
> 
> 
>                   When replying to digests, please change the subject
>         line to
>                   something relevant.
> 
>                   Pawan Kumar wrote:
> 
>                       Hello Justin Sir,
> 
>                       Greetings from Pawan.
>                       Sorry for the late reply.
>                       The max. force was 1.2447973e+o5 on atom 19448.
> 
> 
>                   An Fmax that high is sure to generate problems.  It is
>         up to
>                you to
>                   inspect your system, understand which atoms are
>         interacting
>                to cause
>                   such a force, and determine if you've done something
>         wrong.
> 
>                   -Justin
> 
>                       This particular atom belongs to one of the lipid
>         residues
>                of the
>                       bilayer.
>                       I get Lincs warnings whenever I run the position
>                restraint mdrun.
> 
>                       Thanking you,
>                       Pawan
> 
> 
>                       On Sat, Mar 21, 2009 at 6:08 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:
> 
>                              Hello Justin Sir,
> 
>                              Greetings from Pawan
>                              Thanks for your valuable suggestion and reply.
>                              Initially I gave the emtol of 1000 and the
>         output
>                I got was :
>                              Steepest Descents converged to machine
>         precision
>                in 163 steps
>                              but did not reach the requested Fmax<1000.
>                              Potential Energy = - 4.4516497e+05
> 
> 
>                          ...and how close did Fmax get to 1000?
> 
> 
>                              Even I tried to minimize the popc bilayer
>         which I
>                took from
>                              Tieleman sir's website ( before generating a
>                bigger bilayer
>                              using genconf ) but that also converged to
>         machine
>                       precision but
>                              not to the requested Fmax<1000. How do I
>         proceed
>                further ?
> 
> 
>                          Well, these things are not absolute; Fmax = 1000 is
>                kind of a
>                       rule
>                          of thumb that I use in my own work, but
>         sometimes it
>                is not
>                          necessary.  Careful equilibration should
>         massage your
>                system into
>                          cooperating.
> 
>                          -Justin
> 
>                              Thanks for your suggestions and help.
> 
>                              Thanking you,
>                              Pawan
> 
>                              On Thu, Mar 19, 2009 at 9:13 PM,
> 
>                                 Pawan Kumar wrote:
>                                  > Hello Justin Sir,
>                                  >
>                                  > Greetings from Pawan
>                                  > Thanks for your valuable suggestion
>         and reply.
>                                  > After inserting the protein in the
>         bilayer using
>                       genbox I have
>                                 minimized
>                                  > the whole system without using any
>         position
>                restraints
>                              (i.e. define =
>                                  > -DFLEXIBLE in em.mdp file). I used
>         vanderwaal's
>                       distance
>                              parameter (
>                                  > -vdwd of 0.6 ) in the genbox step.
>                                  > After running mdrun for energy
>         minimization
>                I got the
>                              output as :
>                                  > Steepest Descents converged to
>         Fmax<2250 in
>                14 steps.
>                                  > Potential energy = - 6.9484700e+05
>                                  > Maximum force = 2.2114819e+03 on atom
>         34277
>                                  > Norm. of force = 5.0103039e+04
>                                  >
> 
>                                 You should try for an Fmax of no greater
>         than 1000.
>                        2250 is
>                              still
>                                 very high.
> 
>                                  > I tried decreasing the emtol value in
>         the em.mdp
>                       file but it
>                                 ended with
>                                  > machine precision.
> 
>                                 How far did it converge?  What was Fmax?
> 
>                                  > I have read in literature that 5000
>         steps of
>                Steetest
>                              Descents run is
>                                  > required after inserting the protein
>         in the
>                bilayer
>                       which
>                              should be
>                                  > followed by atleast 1000 steps of
>         conjugate
>                       gradients. How
>                              can I
>                                  > accomplish this ? Is there any
>         parameter to be
>                       given in the
>                                 em.mdp file
>                                  > ? I use steep as the integrator in
>         the mdp
>                file for
>                       energy
>                                 minimization.
> 
>                                 Read the manual.
> 
>                                 Whether or not that exact setup is going
>         to be
>                       "required" is
>                              likely
>                                 system-specific.  I would say that as
>         long as
>                your system
>                              converges
>                                 to a stable,
>                                 negative Epot with a reasonable Fmax
>         (less than
>                1000,
>                       but ideally
>                                 lower) then
>                                 you *may* have an appropriate starting
>         structure.
> 
>                                 -Justin
> 
>                                  > Please help with some suggestions.
>                                  > Thanks in advance.
>                                  >
>                                  > Thanking you,
>                                  > Pawan
>                                  >
>                                  >
>                                  >
> 
> 
> 
>                                        
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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>
>                <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
> 
>            ========================================
> 
> 
> 
>     -- 
>     ========================================
> 
>     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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