[gmx-users] Re: Re: Loss of bonds in HEME iron after pdb2gmx

saradas at ncbs.res.in saradas at ncbs.res.in
Thu Nov 27 06:06:52 CET 2008


Hi Justin,
Thanks a lot for the suggestions regarding the force field and the
sulphide bond. The bond is now formed after changing the distance range in
specbond.dat.
Thanks.
Sarada
>
> Message: 2
> Date: Wed, 26 Nov 2008 07:21:37 -0500
> From: "Justin A. Lemkul" <jalemkul at vt.edu>
> Subject: Re: [gmx-users] Re:  Loss of bonds in HEME iron after pdb2gmx
> To: Discussion list for GROMACS users <gmx-users at gromacs.org>
> Message-ID: <492D3F51.4020708 at vt.edu>
> Content-Type: text/plain; charset=UTF-8; format=flowed
>
>
>
> saradas at ncbs.res.in wrote:
>> Hi Justin,
>> Thanks for your reply. I am using the gmx Gromacs Forcefield. As you
>> said
>> the topology file does contain the bonds and I need not have made the
>> modificatins to specbond.dat. But using the original specbond.dat does
>> not
>> prevent the protonation of CYS. I renamed CYS as CYS2 in the input pdb
>> file and used the following command line
>>         pdb2gmx_mpi -ignh -ff gmx -f 2c9_s50.pdb -o protein_h.pdb -p
>> topology.top -water spce
>> Am I  missing some crucial option here (like -ss for disulphide bonds)?
>> Thanks once again.
>
> One piece of general advice - don't use ffgmx.  It has long been
> deprecated
> (according to the manual, the output of pdb2gmx, and dozens of posts on
> this
> list).  Use one of the newer Gromos96 force fields.
>
> Now, to address the actual question :)  The only thing I can think of is
> that
> specbond.dat uses a simple distance search to determine if the atoms are
> bonded,
> and I believe the tolerance is something like 10%.  So the value in
> specbond.dat
> of 0.25 implies that pdb2gmx should expect the S and Fe atoms to be 0.25
> +/- 0.2
> nm apart.  If they are much further apart, no bond will be detected.
> Check your
> input .pdb file to see how far apart these two atoms are.
>
> -Justin
>
>> Regards,
>> Sarada
>>
>>
>>> Message: 6
>>> Date: Tue, 25 Nov 2008 07:17:14 -0500
>>> From: "Justin A. Lemkul" <jalemkul at vt.edu>
>>> Subject: Re: [gmx-users] Loss of bonds in HEME iron after pdb2gmx
>>> To: Discussion list for GROMACS users <gmx-users at gromacs.org>
>>> Message-ID: <492BECCA.5000708 at vt.edu>
>>> Content-Type: text/plain; charset=UTF-8; format=flowed
>>>
>>>
>>>
>>> saradas at ncbs.res.in wrote:
>>>> Hello,
>>>> Sorry, I realised the HEME HEC naming was not the problem. But still
>>>> the
>>>> pdb2gmx causes loss of all bonds of FE in HEME. The cystine also gets
>>>> protonated and doesnot form a bond with HEME. I tried to preserve the
>>>> bonds by editing the specbond.dat file. I do not know if it can be
>>>> used
>>>> for intramolecular bonding. Kindly inform if the modifications I have
>>>> made
>>>> are valid. The runtime information during the execution of pdb2gmx
>>>> indicated the linking of FE in HEME to the N atoms. However, I do not
>>>> see
>>>> the bonds when I open the output file in Pymol. Please indicate how
>>>> this
>>>> problem can be resolved.
>>>>
>>> Well, using visualization software may not indicate anything.  Most
>>> programs
>>> decide on bonds based on distance, not any sort of intelligent
>>> mechanism.
>>> The
>>> question is whether or not these bonds show up in your topol.top.
>>>
>>>> This is the content of my specbond.dat file:
>>>>
>>>> 5
>>>> CYS     SG      1       HEME    FE      5       0.25    CYS     HEME
>>> Well, this may be why your Cys is getting protonated - you're telling
>>> pdb2gmx
>>> that it should be!  See the format of specbond.dat here:
>>>
>>> http://wiki.gromacs.org/index.php/specbond.dat
>>>
>>> The second-to-last column should be the new residue name for cysteine,
>>> probably
>>> you want something like CYS2.
>>>
>>>> HEME    FE      5       HEME    NA      3       0.22    HEME    HEME
>>>> HEME    FE      5       HEME    NB      3       0.22    HEME    HEME
>>>> HEME    FE      5       HEME    NC      3       0.22    HEME    HEME
>>>> HEME    FE      5       HEME    ND      3       0.22    HEME    HEME
>>>>
>>> Why is this section (above) even necessary?  Are these bonds not
>>> defined
>>> in the
>>> .rtp file of your force field?  Which force field are you using?  I
>>> know
>>> these
>>> are defined in the Gromos-type force fields.
>>>
>>> Try again with the original specbond.dat, but don't assume that
>>> visualization
>>> software will always indicate the presence/absence of a bond.  Examine
>>> your
>>> topology for that.
>>>
>>> -Justin
>>>
>>>> And these I obtained during the execution of pdbgmx:
>>>>
>>>> Opening library file specbond.dat
>>>> 5 out of 5 lines of specbond.dat converted succesfully
>>>> Special Atom Distance matrix:
>>>>                   CYS122  CYS135  CYS143  CYS146  CYS150  CYS237
>>>> CYS309
>>>>                    SG958  SG1059  SG1118  SG1136  SG1165  SG1902
>>>> SG2475
>>>>   CYS135  SG1059   1.677
>>>>   CYS143  SG1118   1.076   1.092
>>>>   CYS146  SG1136   1.313   1.792   0.749
>>>>   CYS150  SG1165   1.175   2.054   0.978   0.426
>>>>   CYS237  SG1902   2.194   3.510   2.536   1.933   1.632
>>>>   CYS309  SG2475   1.745   2.703   2.469   2.507   2.423   2.449
>>>>   CYS343  SG2748   4.679   4.998   4.364   3.763   3.870   3.329
>>>> 4.426
>>>>   CYS406  SG3265   2.258   2.657   2.051   1.603   1.744   1.886
>>>> 2.198
>>>>   CYS457  SG3657   2.232   0.829   1.894   2.582   2.825   4.173
>>>> 2.898
>>>>  HEME462  FE3693   2.257   2.566   1.949   1.489   1.669   1.956
>>>> 2.343
>>>>  HEME462  NA3694   2.456   2.680   2.110   1.663   1.862   2.118
>>>> 2.486
>>>>  HEME462  NB3695   2.199   2.426   1.883   1.498   1.704   2.088
>>>> 2.247
>>>>  HEME462  NC3696   2.061   2.462   1.795   1.321   1.479   1.809
>>>> 2.217
>>>>  HEME462  ND3697   2.331   2.711   2.031   1.505   1.657   1.841
>>>> 2.455
>>>>                   CYS343  CYS406  CYS457 HEME462 HEME462 HEME462
>>>> HEME462
>>>>                   SG2748  SG3265  SG3657  FE3693  NA3694  NB3695
>>>> NC3696
>>>>   CYS406  SG3265   2.501
>>>>   CYS457  SG3657   5.547   3.210
>>>>  HEME462  FE3693   2.512   0.219   3.147
>>>>  HEME462  NA3694   2.361   0.325   3.240   0.211
>>>>  HEME462  NB3695   2.647   0.293   2.977   0.208   0.295
>>>>  HEME462  NC3696   2.671   0.286   3.064   0.209   0.420   0.297
>>>>  HEME462  ND3697   2.389   0.314   3.318   0.208   0.297   0.416
>>>> 0.293
>>>>
>>>>
>>>> Linking HEME-462 FE-3693 and HEME-462 NA-3694...
>>>> Linking HEME-462 FE-3693 and HEME-462 NB-3695...
>>>> Linking HEME-462 FE-3693 and HEME-462 NC-3696...
>>>> Linking HEME-462 FE-3693 and HEME-462 ND-3697...
>>>> N-terminus: PRO-NH2+
>>>> C-terminus: COO-
>>>> Now there are 462 residues with 4749 atoms
>>>> Making bonds...
>>>>
>>>> Thanks.
>>>> Sarada
>>>> Graduate Student,
>>>> NCBS, Bangalore
>>>>
>>>>
>>>>
>>>>
>>>> Hello,
>>>> I am working with the simulation of cytochromeP450. In the output of
>>>> the
>>>> pdb2gmx command, the iron atom in the HEME loses all the bonds,
>>>> including
>>>> the sulphide bond with cystine. Also, the command changed the residue
>>>> name
>>>> from HEME to HEC. During the execution of the pdb2gmx, I get this
>>>> warning:
>>>>
>>>> Warning: 'HEC' not found in residue topology database, trying to use
>>>> 'HEME'
>>>>
>>>> Can someone please tell me, if this is responsible for the loss of
>>>> bonds
>>>> especially the one between CYS and FE. How can the bonds be retained?
>>>> Thanks.
>>>> Sarada
>>>>
>>>>
>>>>
>>>>
>>>>
>>>>
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>>> --
>>> ========================================
>>>
>>> Justin A. Lemkul
>>> Graduate Research Assistant
>>> 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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>>> End of gmx-users Digest, Vol 55, Issue 132
>>> ******************************************
>>>
>>
>>
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>
> --
> ========================================
>
> Justin A. Lemkul
> Graduate Research Assistant
> Department of Biochemistry
> Virginia Tech
> Blacksburg, VA
> jalemkul[at]vt.edu | (540) 231-9080
> http://www.bevanlab.biochem.vt.edu/Pages/Personal/justin
>
> ========================================
>
>
> ------------------------------
>
> Message: 3
> Date: Wed, 26 Nov 2008 07:25:07 -0500
> From: "Justin A. Lemkul" <jalemkul at vt.edu>
> Subject: Re: [gmx-users] Re:  Loss of bonds in HEME iron after pdb2gmx
> To: Discussion list for GROMACS users <gmx-users at gromacs.org>
> Message-ID: <492D4023.3060206 at vt.edu>
> Content-Type: text/plain; charset=UTF-8; format=flowed
>
>
>
> Justin A. Lemkul wrote:
>
>> S and Fe atoms to be 0.25 +/- 0.2 nm apart.  If they are much further
>
> Edit that: 0.25 nm +/- 0.02 nm.  Apparently my math (and/or typing) skills
> are
> suspect first thing in the morning :)
>
> -Justin
>
> --
> ========================================
>
> Justin A. Lemkul
> Graduate Research Assistant
> 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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