[gmx-users] Adding TPO and SEP to G53a6 Forcefield

lalithkumar lalithkumar07 at gmail.com
Sat Feb 22 12:33:32 CET 2014


Dear gromacs users, 

I have got two phosphorylated residues Tyrosine (TPO) and Serine (SEP) in A
and B chains of my heterotrimer homology model. So, I went with the step by
step process suggested by GROMACS at the
link:http://www.gromacs.org/Documentation/How-tos/Adding_a_Residue_to_a_Force_Field

Please dont mind, to be clear, I would like to place each and every single
step that I have made and pasted in ff folder. 

1. I copied a folder named gromos53a6.ff and file residuetypes.dat from the
gromacs installed directory and pasted in the present working directory to
trial the modifications. 

2. I have added topologies for TPO and SEP  to the .rtp file at the end of
file as below: (I am very happy if anyone corrects me if there were any
mistakes) 

[ TPO ] 
 [ atoms ] 
    N     N    -0.31000     0 
    H     H     0.31000     0 
   CA   CH1     0.00000     1 
   CB   CH1     0.00000     2 
  OG1    OA    -0.50000     2 
    P     P     0.90000     2 
  O1P    OP    -0.80000     2 
  O2P    OP    -0.80000     2 
  O3P    OP    -0.80000     2 
  CG2   CH3     0.00000     3 
    C     C       0.450     4 
    O     O      -0.450     4 
 [ bonds ] 
    N     H    gb_2     
    N    CA    gb_20   
   CA     C    gb_26   
    C     O    gb_4     
    C    +N    gb_9     
   CA    CB    gb_26   
   CB   OG1    gb_17   
   CB   CG2    gb_26   
  OG1     P    gb_27     
    P   O1P    gb_23     
    P   O2P    gb_23     
    P   O3P    gb_23   
 [ angles ] 
;  ai    aj    ak   gromos type 
   -C     N     H     ga_31   
    H     N    CA     ga_17   
   -C     N    CA     ga_30   
    N    CA     C     ga_12   
   CA     C    +N     ga_18   
   CA     C     O     ga_29   
    O     C    +N     ga_32   
    N    CA    CB     ga_12   
    C    CA    CB     ga_12   
   CA    CB   OG1     ga_12   
   CA    CB   CG2     ga_14   
  OG1    CB   CG2     ga_14   
   CB   OG1     P     ga_25 
  OG1     P   O1P     ga_13 
  OG1     P   O2P     ga_13 
  OG1     P   O3P     ga_13 
  O1P     P   O2P     ga_28 
  O1P     P   O3P     ga_28 
  O2P     P   O3P     ga_28 
 [ impropers ] 
;  ai    aj    ak    al   gromos type 
    N    -C    CA     H     gi_1     
    C    CA    +N     O     gi_1     
   CA     N     C    CB     gi_2     
   CB   OG1   CG2    CA     gi_2     
 [ dihedrals ] 
;  ai    aj    ak    al   gromos type 
  -CA    -C     N    CA     gd_4     
   -C     N    CA     C     gd_19   
    N    CA     C    +N     gd_20   
    N    CA    CB   OG1     gd_17   
   CA    CB   OG1     P     gd_14   
   CB   OG1     P   O1P     gd_11     

[ SEP ] 
 [ atoms ] 
    N     N    -0.31000     0 
    H     H     0.31000     0 
   CA   CH1     0.00000     1 
   CB   CH2     0.00000     2 
   OG    OA    -0.50000     2 
    P     P     0.90000     2 
  O1P    OP    -0.80000     2 
  O2P    OP    -0.80000     2 
  O3P    OP    -0.80000     2 
    C     C       0.450     3 
    O     O      -0.450     3 
 [ bonds ] 
    N     H    gb_2     
    N    CA    gb_20   
   CA     C    gb_26   
    C     O    gb_4     
    C    +N    gb_9     
   CA    CB    gb_26   
   CB    OG    gb_17   
   OG     P    gb_27     
    P   O1P    gb_23     
    P   O2P    gb_23     
    P   O3P    gb_23   
 [ angles ] 
;  ai    aj    ak   gromos type 
   -C     N     H     ga_31   
    H     N    CA     ga_17   
   -C     N    CA     ga_30   
    N    CA     C     ga_12   
   CA     C    +N     ga_18   
   CA     C     O     ga_29   
    O     C    +N     ga_32   
    N    CA    CB     ga_12   
    C    CA    CB     ga_12   
   CA    CB    OG     ga_12   
   CB    OG     P     ga_25 
   OG     P   O1P     ga_13 
   OG     P   O2P     ga_13 
   OG     P   O3P     ga_13 
  O1P     P   O2P     ga_28 
  O1P     P   O3P     ga_28 
  O2P     P   O3P     ga_28   
 [ impropers ] 
;  ai    aj    ak    al   gromos type 
    N    -C    CA     H     gi_1     
    C    CA    +N     O     gi_1     
   CA     N     C    CB     gi_2     
 [ dihedrals ] 
;  ai    aj    ak    al   gromos type 
  -CA    -C     N    CA     gd_4     
   -C     N    CA     C     gd_19   
    N    CA     C    +N     gd_20   
    N    CA    CB    OG     gd_17   
   CA    CB    OG     P     gd_14   
   CB    OG     P   O1P     gd_11 

3. Created an appropriate entries in .hdb files at the end of file for
hydrogen addition 

TPO 1 
1	1	H       N	-C	CA 
SEP 1 
1	1	H       N	-C	CA 

4. Addition of atom types in .atp file at the end 

   OP  15.99940 ;     phosphate oxygen (P=O) selon hansson aqvist jmb 97 

and in ffnonbonded.itp file added: 

   OP    8      0.000      0.000     A  0.0022153     3.8239e-06 

5. I dont think these are new bonded types and so not disturbed ffbonded.itp
file. 

6. Added TPO and SEP to residuetypes.dat file alphabetically and specifying
it as protein 

7. No special connectivity and so not edited specbond.dat 

After doing these steps, I called the forcefield within working directory
while pdb2gmx prompted. No errors and my file got processed until genbox.
The command issued was: 

g_pdb2gmx -ignh -f protein.pdb -o protein_pdb2gmx.pdb -p
protein_topology.top -water spce 

g_editconf -bt cubic -f protein_pdb2gmx.pdb -o protein_editconf.pdb -d 1.0
-c 

g_genbox -cp protein_editconf.pdb -cs spc216.gro -o protein_genbox.pdb -p
protein_topology.top 

g_grompp -f EM.mdp -c protein_genbox.pdb -p protein_topology.top -o
protein_ions.tpr 

and here, I get the following errors: 

Ignoring obsolete mdp entry 'cpp' 
Generated 222 of the 1653 non-bonded parameter combinations 

ERROR 1 [file AMPK_topology_Protein_chain_A.itp, line 15435]: 
  No default LJ-14 types 


ERROR 2 [file AMPK_topology_Protein_chain_A.itp, line 15436]: 
  No default LJ-14 types 


ERROR 3 [file AMPK_topology_Protein_chain_A.itp, line 15437]: 
  No default LJ-14 types 


ERROR 4 [file AMPK_topology_Protein_chain_B.itp, line 7610]: 
  No default LJ-14 types 


ERROR 5 [file AMPK_topology_Protein_chain_B.itp, line 7611]: 
  No default LJ-14 types 


ERROR 6 [file AMPK_topology_Protein_chain_B.itp, line 7612]: 
  No default LJ-14 types 



I am not quite where my bond types went wrong, these lines in my .itp files
are corresponding to atoms listed below: 
15435 1923 1926 
15436 1923 1927 
15437 1923 1928 

ATOM   1920  N   TPO A 183      68.232  63.098  68.923  1.00  0.00             
ATOM   1921  H   TPO A 183      68.988  63.061  69.576  1.00  0.00             
ATOM   1922  CA  TPO A 183      67.034  62.439  69.431  1.00  0.00             
ATOM   1923  CB  TPO A 183      67.071  62.360  70.916  1.00  0.00             
ATOM   1924  OG1 TPO A 183      68.205  61.626  71.328  1.00  0.00             
ATOM   1925  P   TPO A 183      69.170  62.266  72.397  1.00  0.00             
ATOM   1926  O1P TPO A 183      70.456  61.480  72.420  1.00  0.00             
ATOM   1927  O2P TPO A 183      68.524  62.221  73.762  1.00  0.00             
ATOM   1928  O3P TPO A 183      69.457  63.705  72.029  1.00  0.00             
ATOM   1929  CG2 TPO A 183      65.822  61.647  71.391  1.00  0.00             
ATOM   1930  C   TPO A 183      65.781  63.123  68.984  1.00  0.00             
ATOM   1931  O   TPO A 183      65.536  64.269  69.348  1.00  0.00         

which states CB connected phosphate group oxygens, if i am correct... but
how it happened is i dont know 

similar is the case with next 3 errors that I have not added here. 

I request if anyone could highlight the errors with my procedure and provide
a needful information to rectify it. Also, I am excited to know, Is there
any chance, to get g53a6ff prepared files for phosphorylated residues.
please, let me know for any further information 

Many thanks in advance 

sincerely, 
Lalith Kumar K 
Research Associate, 
University of Hyderabad Campus 
India 

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