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Re: [modeller_usage] Protein + Metal Ion



Modeller Caretaker wrote:

<">mailto:>

Anshul Awasthi wrote:

I am trying to model the structure of a protein that contains Zn ion in the active site, using modeller 7v7. For the modeling purpose I am using 5 templates and one of them contains 2 His residues coordinated with Zn ion, and these residues are conserved among all the templates and the target sequence.



I suggest that you use the latest version, Modeller 8v2, instead. Not only does it incorporate a number of bug fixes, but it gives more informative diagnostics for problems like this one. It will read old TOP scripts just fine, provided they have the .top file extension.

I have put /z* at the end of the alignment file as follows:

P1;1WVR
structureX:1WVR:   9 :A: 181 :::::
PRKPEIQNEIIDLHNSLRRSVN------------PTASNMLKMEWYPEAAANAERWAYRCIESHSSRDSRVIGG- ---IKCGENIYMAT-----YPAKWTDIIHAWHGEYKDFKYGVGAVPSD----AVIGHYTQIVWYKSYRAGCAAAY
CPSSKYS------YFYVCQYCPAGNIIGKTATPYKSGPPCGDCP-SD-C-DN--GLCTN/z/*



The final / is unnecessary here, since nothing follows the ion.

==================================================================
But when I run the modeller i get an error message stating "SEQUENCE DIFFERENCE BETWEEN ALIGNMENT AND PDB "

rdabrk__291E> Sequence difference between alignment and  pdb :

STRUCTURE RES_IND  ALN_ITYP  ALN_RES  X_ITYP  X_RES  -----*-----
2 174 30 ZN2 24 UNK GLCTNz



Your PDB file appears to have an UNK residue rather than a Zn atom here. It's hard to say what your problem might be, as I'm guessing your PDB is modified from code 1wvr (since the A chain continues to residue 221, and the ion is cadmium, not zinc).

If your problem continues, send me your full input files, and I'll take a look.

    Ben Webb, Modeller Caretaker


Dear Ben,
I was able to run the programme and got a model with Zn ion. But I was only able to do it without adding the restraints to the His residues and the Zn ion. But when I put the following subroutine at the end of my .top file, I get an error.

SUBROUTINE ROUTINE        = 'special_restraints'
SET ATOM_IDS 'NE2:420' 'ZN2:1436'
ADD_RESTRAINT RESTRAINT_PARAMETERS = 3 1 1 27 2 2 0 2.2 0.1
SET ATOM_IDS 'NE2:892' 'ZN2:1436'
ADD_RESTRAINT RESTRAINT_PARAMETERS = 3 1 1 27 2 2 0 2.2 0.1
RETURN
END_SUBROUTINE

Following is the last portion of the log file with the error;

TOP_________>   231  199 MAKE_RESTRAINTS

make_re_417_> Restraint type to be calculated:  distance
make_re_422_> Number of previous, current restraints : 19104 19104 make_re_423_> Number of previous, current selected restraints: 18913 18913
TOP_________>   232  200 DELETE_ALIGNMENT

TOP_________>   233  201 RETURN

TOP_________>   234  165 CALL ROUTINE = 'special_restraints'

TOP_________>   235  810 SET ATOM_IDS 'NE2:420' 'ZN2:1436'

TOP_________> 236 811 ADD_RESTRAINT RESTRAINT_PARAMETERS = 3 1 1 27 2 2 0 2.2 0;
                    .1

add_res_442E> One or more atoms absent from MODEL:  NE2:420 ZN2:1436
top_error__E> ERROR_STATUS >= STOP_ON_ERROR:        1       1

The number of the residues I have used correspond to the modeller numbering system (*.ini file). I am not sure about the RESTRAINT_PARAMETERS I have put in the subroutines as I have duplicated it from one of the earlier posts. I tried to know more about them from the manual but could not understand. I shall be glad if you could also explain the meaning of various fields being used in the parameters.

With regards,
Anshul