January 2006 - modeller_usage

Modeling antibody
by BiplabBose＠bits-pilani.ac.in 09 Jan '06

09 Jan '06

Hi, I am trying to model an antibody variable region using Modeller. I have two questions : 1. I have modelled the heavy chain and Light chain seperately. Now, how can I orient this two chains to generate the complete molecule? 2. CDR loops of antibodies have specific canonical structures, which depends upon loop length, hydrogen bond ladder etc. Can I modify Modeller to build the loops as per the canonical structures? With regards Biplab -- Biplab Bose, Ph. D Lecturer, Biological Sciences Group, Birla Institute of Technology & Science (BITS), Pilani, Rajasthan, INDIA PIN-333031 Ph.# 9828717874 http://www.biplabbose.wb.st http://www.bits-pilani.ac.inBITS, Pilani

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varied models
by Inbal Halperin 06 Jan '06

06 Jan '06

Hi fellow Modellers ! Does anyone know how can I create variable models? I want to get models that are different from one another (high rmsd between models). Thanks ! -Inbal

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varied models
by Inbal Halperin 06 Jan '06

06 Jan '06

Hi fellow Modellers ! Does anyone know how can I create variable models? I want to get models that are different from one another (high rmsd between models). Thanks ! -Inbal

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Re: Ligand clues and water blues
by Mohsen, Al-Walid 06 Jan '06

06 Jan '06

Hi Jo, You have two messages and I am not sure where you are. I mentioned earlier in my reply to replace the /...* with XXX* for the template and ...* for the model in your alignment file. It seems like you're still including the '/' character, which is unnecessary at the end of the sequence and may cause problems (shifts). The '/' is necessary to break the peptide sequence in the middle of the sequence, not at the end. IF you do a dimer for example you need it there between the monomers, but NOT at the end. In option one, > this gives this error: > rdabrk__290E> Number of residues in the alignment and pdb files are > different: 517 516 > For alignment entry: 1 > recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 the error is real and you should look for where the problem is in the pdb file vs the alignment file. As I understand it you should have the residues coordinates in the pdb file then two ligands AMP (515) and Mg (516), and then one water molecule, which should be marked as HETATM with the number 517. If this is the way it is and the second line of the template in alignment file is: structureX:Bamu:1 : :517H : :unknown:unknown:-1.00:-1.00 RKQLPEPDLTFXXX* and the XXX* is aligned with the ...* AFTER the .ini is written (see below), then you should not get this error, unless you deleted something (a residue) by mistake. In option 2: You MUST check the alignment file for shifts after you exe or start the Modeler_Run AND the .ini file is written. Modeler 7 is known for introducing '-' into the sequence and things get messed up. This occurs near the '/' character. In option 3: Everyone else can correct me, if I am wrong, but I do not see the problem with water coming up as BLK. As long as it's in the proper position in the 3D structure, it's OK. Just replace BLK with HOH manually in the pdb file. If you do the XXX aligned with ... then all HETATM including water are treated as HETATM and renamed BLK. In option 4: I would leave water_IO = on out. Do not assume that all the commands work perfectly right. As I mentioned above Modeler 7 has this shift issues that was not present in Modeler 6. Best regards, Walid Mohsen > From: <modeller_usage-request(a)salilab.org> > Reply-To: <modeller_usage(a)salilab.org> > Date: Fri, 6 Jan 2006 04:21:40 -0800 > To: <modeller_usage(a)salilab.org> > Subject: modeller_usage Digest, Vol 5, Issue 5 > > Send modeller_usage mailing list submissions to > modeller_usage(a)salilab.org > > To subscribe or unsubscribe via the World Wide Web, visit > http://salilab.org/mailman/listinfo/modeller_usage > or, via email, send a message with subject or body 'help' to > modeller_usage-request(a)salilab.org > > You can reach the person managing the list at > modeller_usage-owner(a)salilab.org > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of modeller_usage digest..." > > > Today's Topics: > > 1. modelling with water again (jo hanna) > 2. Help needed (jo hanna) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Wed, 4 Jan 2006 00:33:28 +0000 > From: jo hanna <jfhanna(a)gmail.com> > Subject: [modeller_usage] modelling with water again > To: modeller_usage(a)salilab.org > Message-ID: > <4b21263d0601031633p52f2b0dbid0d1190a1301f952(a)mail.gmail.com> > Content-Type: text/plain; charset="iso-8859-1" > > Hello > > Many thanks for the advice Walid, I have done as you have suggested and have > made some progress but still am not seeing the behaviour that I am > expecting. > > I have been using the pap format for the alignment as this has being working > better and have tried the following varied options in the top and align > files, I should also mention that in my pdb i have the exact number of > ligands(2), and single water that i would want to include in my model > > Option 1: > remove water_io command from top file, > try X for water and not w therefore at end of alignment > structure = /..W > and seqeunce = /... > > this gives this error: > rdabrk__290E> Number of residues in the alignment and pdb files are > different: 517 516 > For alignment entry: 1 > recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 > > Option 2: > water_io = on in top file, in alignment file, structure end aligmment = /..X > seqeunce end alignment = /... > gives this error > rdabrk__291E> Sequence difference between alignment and pdb : > > STRUCTURE RES_IND ALN_ITYP ALN_RES X_ITYP X_RES -----*----- > 1 517 24 UNK 28 TIP3 LTF..X > check_a_337E> Structure not read in: 1 > recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 > > Option 3: > water_io = on in top file, end of alignments, strcuture = /..w seqeunce = > /... > > This combination creates a model but there is no mention in the logfile of > the BLK residues or of whether the water is accepted and in the final model > the water O is written as a BLK residue. From previoud posts i was expecting > a message in the log file re the TIP3 water and also for the model to > contain a HOH or OH2 and not a BLK. > > Option 4: > water_io = on in top file and end os alignments strcuture = /..w seqeunce = > /..w > This once agaian produces the out of range error > delete__443_> Restraints marked for deletion were removed. > Total number of restraints before, now: 51247 47550 > nselat__596E> Atom index is out of range (iatm,natm): 4033 4032 > recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 > > Any help would be greatly apprecaited. > > Thanks > Jo >

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Help needed
by jo hanna 06 Jan '06

06 Jan '06

Hello I have been trying for some time now to model with 2 ligands and 1 water molecule and I feel that I have tried everything from the list, faq and tutorials and not got very far. I have posted a number of questions to the list but can still not get this to work, properly! I have managed to include the ligands and the water O into my output pdb by using the script and alignment below, however there is not recognition that this oxygen is a water and in the output model it is listed as BLK along with the other ligands. From reading posts in the user list I was expecting this water oxygen to be recognised as such, since i have used the w character to represent it and also HOH in the pdb file (the file has been edited to include only the protain, ligand and water that I am wishing to model around). I cannot type w at the end of the sequence alignment however as this causes modeller to fail with an out of range error. I would really appreciate some help as I have completelty exhaused all of my options now. Thanks Jo top INCLUDE SET ALNFILE = 'CcHh1-Bamu.pap', ALIGNMENT_FORMAT = 'PAP' SET KNOWNS = 'Bamu' SET SEQUENCE = 'CcHh1' SET STARTING_MODEL = 1 SET ENDING_MODEL = 1 SET HETATM_IO = on, WATER_IO = on CALL ROUTINE = 'model' end of alignment (pap format) alignment _aln.pos 480 490 500 510 520 Bamu RKQLPEPDLTF/..w CcHh1 RNALPEPVIPA/... produce model with water included as: HETATM 4032 O BLK 525 37.988 89.184 33.629 1.00 25.09 2SG4034 END

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modelling with water again
by jo hanna 03 Jan '06

03 Jan '06

Hello Many thanks for the advice Walid, I have done as you have suggested and have made some progress but still am not seeing the behaviour that I am expecting. I have been using the pap format for the alignment as this has being working better and have tried the following varied options in the top and align files, I should also mention that in my pdb i have the exact number of ligands(2), and single water that i would want to include in my model Option 1: remove water_io command from top file, try X for water and not w therefore at end of alignment structure = /..W and seqeunce = /... this gives this error: rdabrk__290E> Number of residues in the alignment and pdb files are different: 517 516 For alignment entry: 1 recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 Option 2: water_io = on in top file, in alignment file, structure end aligmment = /..X seqeunce end alignment = /... gives this error rdabrk__291E> Sequence difference between alignment and pdb : STRUCTURE RES_IND ALN_ITYP ALN_RES X_ITYP X_RES -----*----- 1 517 24 UNK 28 TIP3 LTF..X check_a_337E> Structure not read in: 1 recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 Option 3: water_io = on in top file, end of alignments, strcuture = /..w seqeunce = /... This combination creates a model but there is no mention in the logfile of the BLK residues or of whether the water is accepted and in the final model the water O is written as a BLK residue. From previoud posts i was expecting a message in the log file re the TIP3 water and also for the model to contain a HOH or OH2 and not a BLK. Option 4: water_io = on in top file and end os alignments strcuture = /..w seqeunce = /..w This once agaian produces the out of range error delete__443_> Restraints marked for deletion were removed. Total number of restraints before, now: 51247 47550 nselat__596E> Atom index is out of range (iatm,natm): 4033 4032 recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 Any help would be greatly apprecaited. Thanks Jo

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Re: Adding water molecules as ligands (minor corrections); modeller_usage Digest, Vol 5, Issue 3
by Mohsen, Al-Walid 03 Jan '06

03 Jan '06

Please note the corrections below: Best regards, Walid > > Today's Topics: > > 1. Re: modeller_usage Digest, Vol 5, Issue 2 (Mohsen, Al-Walid) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Tue, 03 Jan 2006 11:09:35 -0500 > From: "Mohsen, Al-Walid" <mohsaw(a)chp.edu> > Subject: [modeller_usage] Re: modeller_usage Digest, Vol 5, Issue 2 > To: <modeller_usage(a)salilab.org> > Message-ID: <BFE00BEF.2014%mohsaw(a)chp.edu> > Content-Type: text/plain; charset="US-ASCII" > > If you're only trying to include 3 water molecules that you know are > important, try the following: > > 1- Edit the pdb file of your template to only include these specific ones, > i.e, delete all other water molecules. Number these three water molecules 515 > 516, and 517. > 2- Edit your .top file deleting the WATER_IO = on command and leave the > HETATM_IO = on command. > 3- I usually use .pir and not .ali but this may still be applicable. Edit > your .ali file as follows: > (a) Change the second line in the paragraph so it will be: > structureX:Bamu:1 : :517H : :unknown:unknown:-1.00:-1.00 > > (b) Remove the /w* at the end of the molecule from both template and > your sequence (model) lines. > (c) After the last residue in the template, add XXX* that would > correspond to the 3 water molecules in the pdb file. > (d) After the last residue in the Sequence (model) lines add ...* > (e) Make sure that the XXX* in the template and the model ...* align exactly > right. It seems you will not have a problem with this since the alignment is > rather simple. > (f) Change the second line in the template paragraph to: > sequence:CcHh1:1 : :523 : :unknown:unknown:-1.00:-1.00 > > I might be wrong but it seems like Modeler is confused about what is "w". > > I hope it works!! > > Best regards, > > Walid Mohsen > > Al-Walid A. Mohsen, Ph.D. > Research Assistant Professor > Department of Pediatrics > School of Medicine > University of Pittsburgh > 3460 Fifth Ave, Rangos 2113 > Pittsburgh, PA 15213 > U.S.A. > Phone: (412) 692-7928 > > > >> From: <modeller_usage-request(a)salilab.org> >> Reply-To: <modeller_usage(a)salilab.org> >> Date: Tue, 3 Jan 2006 02:52:59 -0800 >> To: <modeller_usage(a)salilab.org> >> Subject: modeller_usage Digest, Vol 5, Issue 2 >> >> Send modeller_usage mailing list submissions to >> modeller_usage(a)salilab.org >> >> To subscribe or unsubscribe via the World Wide Web, visit >> http://salilab.org/mailman/listinfo/modeller_usage >> or, via email, send a message with subject or body 'help' to >> modeller_usage-request(a)salilab.org >> >> You can reach the person managing the list at >> modeller_usage-owner(a)salilab.org >> >> When replying, please edit your Subject line so it is more specific >> than "Re: Contents of modeller_usage digest..." >> >> >> Today's Topics: >> >> 1. modelling with ligands again! mainly water issues (jo hanna) >> 2. Modelling with ligands - Not Working! (Joanne Hanna) >> >> >> ---------------------------------------------------------------------- >> >> Message: 1 >> Date: Tue, 3 Jan 2006 10:21:12 +0000 >> From: jo hanna <jfhanna(a)gmail.com> >> Subject: [modeller_usage] modelling with ligands again! mainly water >> issues >> To: modeller_usage(a)salilab.org >> Message-ID: >> <4b21263d0601030221y59eef18eo9a8fc712b6fe89d6(a)mail.gmail.com> >> Content-Type: text/plain; charset="iso-8859-1" >> >> Hello again >> >> In follow up to my own question I have found that the issue only really >> occurs when i try to include the water, either with the other ligand or even >> without (after re ordering of the pdb file) Does the water in the pdb file >> have to have H atoms attached or is a crystallographic O sufficient? >> >> My log with the hetatms only no water, is: >> delete__442E> One or more atoms absent from MODEL: O: 522: C: 522: N: 523: >> CA: 523: >> delete__442E> One or more atoms absent from MODEL: O: 523: C: 523: N: 524: >> CA: 524: >> delete__443_> Restraints marked for deletion were removed. >> Total number of restraints before, now: 51172 47475 >> >> which still shows these atoms absent errors which i don't understand. >> >> When adding just the water and no other ligand my top and aln files are >> appended and my log error is an out of bounds as before. SO it aapears that >> the water is the problem, I have also tried in my log file with only the >> water changing the line SET HETATM_IO = on, WATER_IO = on to just SET >> WATER_IO = on but this maks no difference. >> >> delete__442E> One or more atoms absent from MODEL: O: 522: C: 522: N: 523: >> CA: 523: >> delete__443_> Restraints marked for deletion were removed. >> Total number of restraints before, now: 48782 45085 >> nselat__596E> Atom index is out of range (iatm,natm): 4009 4008 >> recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 >> >> TOP >> INCLUDE >> SET ALNFILE = 'CcHh1-Bamu.ali' >> SET KNOWNS = 'Bamu' >> SET SEQUENCE = 'CcHh1' >> SET STARTING_MODEL = 1 >> SET ENDING_MODEL = 1 >> SET HETATM_IO = on, WATER_IO = on >> CALL ROUTINE = 'model'\ >> >> ALIGNMENT >>> P1;Bamu >> structureX:Bamu:1::515::undefined:undefined:-1.00:-1.00 >> GTHEEEQYLFAVNNTKAEYPRDKTIHQLFEEQVSKRPNNVAIVCENEQLTYHELNVKANQLARIFIEKGIGKDTL >> VGIMMEKSIDLFIGILAVLKAGGAYVPIDIEYPKERIQYILDDSQARMLLTQKHLVHLIHNIQFNGQVEIFEEDT >> IKIREGTNLHVPSKSTDLAYVIYTSGTTGNPKGTMLEHKGISNLKVFFEN-SLNVTEKDRIGQFASISFDASVWE >> MFMALLTGASLYIILKDTINDFVKFEQYINQKEITVITLPPTYVVHL-----DPERILSIQTLITAGSATSPSLV >> NKWKE--KVTYINAYGPTETTICATTWVATKETIGHSVPIGAPIQNTQIYIVDENLQLKSVGEAGELCIGGEGLA >> RGYWKRPELTSQKFVDNPFVP-GEKLYKTGDQARWLSDGNIEYLGRIDNQVKIRGHRVELEEVESILLKHMYISE >> TAVSVHKDHQEQPYLCAYFVSE---KHIPLEQLRQFSSEELPTYMIPSYFIQLDKMPLTSNGKIDRKQLPEPDLT >> F/w* >> >>> P1;CcHh1 >> sequence:CcHh1:1::523::::0.00: 0.00 >> GPAERLELLAGWNDTDADRAR-PGLVEMFREQARARPDAVAVVDERRTLTYRQVEKLSAQLAHQLLARGLAVEDV >> VGISLERSAEMVIGLLAVLRAGGAFVPLDPHWPAERRAVVIEDAGVVVQLDASGEPAPG---EPEAVAVDLGDWR >> FGDRPTEDTGVTVPGDALAYVIFTSGSTGRPKGAMIRHEAISERLLWQIHEILGFGHDDASLFKAPLSFDISINE >> IFLPLVSGGRLVVLRPGGERDPHHLLSVIDEQRVTFTYLVSSMLDVLLEMAGDSGRLDSLRHVWCGGEVLTPELY >> ERFRTRLDIPLYHGYGPAETTIGVSHVVYRGAAERLSTSIGRANPNTRLYVLDDELRPVPVGVGGELYAGGLLLG >> RGYVNAPGLTASRFVANPFADDGSRLYRTGDLARFAPDGSLDFLGRADNQVKIRGMRLEIEDVEVGLAEHPRVRH >> TCVVARKNTAGGTYLVGYVIPAAGHEDLRADEVKAWAGEHMVEYMVPTHVVVMTEFPLTANGKLDRNALPEPVIP >> A/w* >> >> Sorry to post so many mails, I'm really struggling to figure this out! >> >> Jo >>

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Re: modeller_usage Digest, Vol 5, Issue 2
by Mohsen, Al-Walid 03 Jan '06

03 Jan '06

If you're only trying to include 3 water molecules that you know are important, try the following: 1- Edit the pdb file of your template to only include these specific ones, i.e, delete all other water molecules. Number these two water molecules 515 516, and 517. 2- Edit your .top file deleting the WATER_IO = on command and leave the HETATM_IO = on command. 3- I usually use .pir and not .ali but this may still be applicable. Edit your .ali file as follows: (a) Change the second line in the paragraph so it will be: structureX:Bamu:1 : :517H : :unknown:unknown:-1.00:-1.00 (b) Remove the /w* at the end of the molecule from both template and your sequence lines. (c) After the last residue in the template, add XXX* that would correspond to the 3 water molecules in the pdb file. (d) After the last residue in the Sequence lines add ...* (e) Make sure that the XXX* in the template and the ...* align exactly right. It seems you will not have a problem with this since the alignment is rather simple. (f) Change the second line in the template paragraph to: sequence:CcHh1:1 : :523 : :unknown:unknown:-1.00:-1.00 I might be wrong but it seems like Modeler is confused about what is "w". I hope it works!! Best regards, Walid Mohsen Al-Walid A. Mohsen, Ph.D. Research Assistant Professor Department of Pediatrics School of Medicine University of Pittsburgh 3460 Fifth Ave, Rangos 2113 Pittsburgh, PA 15213 U.S.A. Phone: (412) 692-7928 > From: <modeller_usage-request(a)salilab.org> > Reply-To: <modeller_usage(a)salilab.org> > Date: Tue, 3 Jan 2006 02:52:59 -0800 > To: <modeller_usage(a)salilab.org> > Subject: modeller_usage Digest, Vol 5, Issue 2 > > Send modeller_usage mailing list submissions to > modeller_usage(a)salilab.org > > To subscribe or unsubscribe via the World Wide Web, visit > http://salilab.org/mailman/listinfo/modeller_usage > or, via email, send a message with subject or body 'help' to > modeller_usage-request(a)salilab.org > > You can reach the person managing the list at > modeller_usage-owner(a)salilab.org > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of modeller_usage digest..." > > > Today's Topics: > > 1. modelling with ligands again! mainly water issues (jo hanna) > 2. Modelling with ligands - Not Working! (Joanne Hanna) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Tue, 3 Jan 2006 10:21:12 +0000 > From: jo hanna <jfhanna(a)gmail.com> > Subject: [modeller_usage] modelling with ligands again! mainly water > issues > To: modeller_usage(a)salilab.org > Message-ID: > <4b21263d0601030221y59eef18eo9a8fc712b6fe89d6(a)mail.gmail.com> > Content-Type: text/plain; charset="iso-8859-1" > > Hello again > > In follow up to my own question I have found that the issue only really > occurs when i try to include the water, either with the other ligand or even > without (after re ordering of the pdb file) Does the water in the pdb file > have to have H atoms attached or is a crystallographic O sufficient? > > My log with the hetatms only no water, is: > delete__442E> One or more atoms absent from MODEL: O: 522: C: 522: N: 523: > CA: 523: > delete__442E> One or more atoms absent from MODEL: O: 523: C: 523: N: 524: > CA: 524: > delete__443_> Restraints marked for deletion were removed. > Total number of restraints before, now: 51172 47475 > > which still shows these atoms absent errors which i don't understand. > > When adding just the water and no other ligand my top and aln files are > appended and my log error is an out of bounds as before. SO it aapears that > the water is the problem, I have also tried in my log file with only the > water changing the line SET HETATM_IO = on, WATER_IO = on to just SET > WATER_IO = on but this maks no difference. > > delete__442E> One or more atoms absent from MODEL: O: 522: C: 522: N: 523: > CA: 523: > delete__443_> Restraints marked for deletion were removed. > Total number of restraints before, now: 48782 45085 > nselat__596E> Atom index is out of range (iatm,natm): 4009 4008 > recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 > > TOP > INCLUDE > SET ALNFILE = 'CcHh1-Bamu.ali' > SET KNOWNS = 'Bamu' > SET SEQUENCE = 'CcHh1' > SET STARTING_MODEL = 1 > SET ENDING_MODEL = 1 > SET HETATM_IO = on, WATER_IO = on > CALL ROUTINE = 'model'\ > > ALIGNMENT >> P1;Bamu > structureX:Bamu:1::515::undefined:undefined:-1.00:-1.00 > GTHEEEQYLFAVNNTKAEYPRDKTIHQLFEEQVSKRPNNVAIVCENEQLTYHELNVKANQLARIFIEKGIGKDTL > VGIMMEKSIDLFIGILAVLKAGGAYVPIDIEYPKERIQYILDDSQARMLLTQKHLVHLIHNIQFNGQVEIFEEDT > IKIREGTNLHVPSKSTDLAYVIYTSGTTGNPKGTMLEHKGISNLKVFFEN-SLNVTEKDRIGQFASISFDASVWE > MFMALLTGASLYIILKDTINDFVKFEQYINQKEITVITLPPTYVVHL-----DPERILSIQTLITAGSATSPSLV > NKWKE--KVTYINAYGPTETTICATTWVATKETIGHSVPIGAPIQNTQIYIVDENLQLKSVGEAGELCIGGEGLA > RGYWKRPELTSQKFVDNPFVP-GEKLYKTGDQARWLSDGNIEYLGRIDNQVKIRGHRVELEEVESILLKHMYISE > TAVSVHKDHQEQPYLCAYFVSE---KHIPLEQLRQFSSEELPTYMIPSYFIQLDKMPLTSNGKIDRKQLPEPDLT > F/w* > >> P1;CcHh1 > sequence:CcHh1:1::523::::0.00: 0.00 > GPAERLELLAGWNDTDADRAR-PGLVEMFREQARARPDAVAVVDERRTLTYRQVEKLSAQLAHQLLARGLAVEDV > VGISLERSAEMVIGLLAVLRAGGAFVPLDPHWPAERRAVVIEDAGVVVQLDASGEPAPG---EPEAVAVDLGDWR > FGDRPTEDTGVTVPGDALAYVIFTSGSTGRPKGAMIRHEAISERLLWQIHEILGFGHDDASLFKAPLSFDISINE > IFLPLVSGGRLVVLRPGGERDPHHLLSVIDEQRVTFTYLVSSMLDVLLEMAGDSGRLDSLRHVWCGGEVLTPELY > ERFRTRLDIPLYHGYGPAETTIGVSHVVYRGAAERLSTSIGRANPNTRLYVLDDELRPVPVGVGGELYAGGLLLG > RGYVNAPGLTASRFVANPFADDGSRLYRTGDLARFAPDGSLDFLGRADNQVKIRGMRLEIEDVEVGLAEHPRVRH > TCVVARKNTAGGTYLVGYVIPAAGHEDLRADEVKAWAGEHMVEYMVPTHVVVMTEFPLTANGKLDRNALPEPVIP > A/w* > > Sorry to post so many mails, I'm really struggling to figure this out! > > Jo >

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modelling with ligands again! mainly water issues
by jo hanna 03 Jan '06

03 Jan '06

Hello again In follow up to my own question I have found that the issue only really occurs when i try to include the water, either with the other ligand or even without (after re ordering of the pdb file) Does the water in the pdb file have to have H atoms attached or is a crystallographic O sufficient? My log with the hetatms only no water, is: delete__442E> One or more atoms absent from MODEL: O: 522: C: 522: N: 523: CA: 523: delete__442E> One or more atoms absent from MODEL: O: 523: C: 523: N: 524: CA: 524: delete__443_> Restraints marked for deletion were removed. Total number of restraints before, now: 51172 47475 which still shows these atoms absent errors which i don't understand. When adding just the water and no other ligand my top and aln files are appended and my log error is an out of bounds as before. SO it aapears that the water is the problem, I have also tried in my log file with only the water changing the line SET HETATM_IO = on, WATER_IO = on to just SET WATER_IO = on but this maks no difference. delete__442E> One or more atoms absent from MODEL: O: 522: C: 522: N: 523: CA: 523: delete__443_> Restraints marked for deletion were removed. Total number of restraints before, now: 48782 45085 nselat__596E> Atom index is out of range (iatm,natm): 4009 4008 recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 TOP INCLUDE SET ALNFILE = 'CcHh1-Bamu.ali' SET KNOWNS = 'Bamu' SET SEQUENCE = 'CcHh1' SET STARTING_MODEL = 1 SET ENDING_MODEL = 1 SET HETATM_IO = on, WATER_IO = on CALL ROUTINE = 'model'\ ALIGNMENT >P1;Bamu structureX:Bamu:1::515::undefined:undefined:-1.00:-1.00 GTHEEEQYLFAVNNTKAEYPRDKTIHQLFEEQVSKRPNNVAIVCENEQLTYHELNVKANQLARIFIEKGIGKDTL VGIMMEKSIDLFIGILAVLKAGGAYVPIDIEYPKERIQYILDDSQARMLLTQKHLVHLIHNIQFNGQVEIFEEDT IKIREGTNLHVPSKSTDLAYVIYTSGTTGNPKGTMLEHKGISNLKVFFEN-SLNVTEKDRIGQFASISFDASVWE MFMALLTGASLYIILKDTINDFVKFEQYINQKEITVITLPPTYVVHL-----DPERILSIQTLITAGSATSPSLV NKWKE--KVTYINAYGPTETTICATTWVATKETIGHSVPIGAPIQNTQIYIVDENLQLKSVGEAGELCIGGEGLA RGYWKRPELTSQKFVDNPFVP-GEKLYKTGDQARWLSDGNIEYLGRIDNQVKIRGHRVELEEVESILLKHMYISE TAVSVHKDHQEQPYLCAYFVSE---KHIPLEQLRQFSSEELPTYMIPSYFIQLDKMPLTSNGKIDRKQLPEPDLT F/w* >P1;CcHh1 sequence:CcHh1:1::523::::0.00: 0.00 GPAERLELLAGWNDTDADRAR-PGLVEMFREQARARPDAVAVVDERRTLTYRQVEKLSAQLAHQLLARGLAVEDV VGISLERSAEMVIGLLAVLRAGGAFVPLDPHWPAERRAVVIEDAGVVVQLDASGEPAPG---EPEAVAVDLGDWR FGDRPTEDTGVTVPGDALAYVIFTSGSTGRPKGAMIRHEAISERLLWQIHEILGFGHDDASLFKAPLSFDISINE IFLPLVSGGRLVVLRPGGERDPHHLLSVIDEQRVTFTYLVSSMLDVLLEMAGDSGRLDSLRHVWCGGEVLTPELY ERFRTRLDIPLYHGYGPAETTIGVSHVVYRGAAERLSTSIGRANPNTRLYVLDDELRPVPVGVGGELYAGGLLLG RGYVNAPGLTASRFVANPFADDGSRLYRTGDLARFAPDGSLDFLGRADNQVKIRGMRLEIEDVEVGLAEHPRVRH TCVVARKNTAGGTYLVGYVIPAAGHEDLRADEVKAWAGEHMVEYMVPTHVVVMTEFPLTANGKLDRNALPEPVIP A/w* Sorry to post so many mails, I'm really struggling to figure this out! Jo

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Problems with modelling with ligand
by jo hanna 03 Jan '06

03 Jan '06

Hello I apologise if this message is posted more than once, I am having difficulties with my email system. My questions regards the process for modelling with ligands. >From the tutorial i understand that you have the ligands in the pdb file template and then you manually alter the alignment to take account of these ligands, for me that means adding /..w* to the end of the structure and seqeunce alignments (i'm unsure if you add it to the seqeunce, if you want it included). This also means changing the number of residues in the structure file in the alignment to include the 3 BLK/water items, as even if they are included in the pdb prior to alignment they are not included in the residue number line in the alignment (i.e. in alignment below changing original 514 to 517). I then do the modelling using the alignment, as appended below and the model.top script (7v7 user!) also below and get the following error which i do not understand. check_ali___> Checking the sequence-structure alignment. Implied target CA(i)-CA(i+1) distances longer than 8.0 angstroms: ALN_POS TMPL RID1 RID2 NAM1 NAM2 DIST ---------------------------------------------- 134 1 134 138 G E 8.407 END OF TABLE delete__442E> One or more atoms absent from MODEL: O: 522: C: 522: N: 523: CA: 523: delete__442E> One or more atoms absent from MODEL: O: 523: C: 523: N: 524: CA: 524: delete__442E> One or more atoms absent from MODEL: O: 524: C: 524: N: 525: CA: 525: delete__443_> Restraints marked for deletion were removed. Total number of restraints before, now: 51247 47550 nselat__596E> Atom index is out of range (iatm,natm): 4033 4032 recover____E> ERROR_STATUS >= STOP_ON_ERROR: 1 1 Dynamically allocated memory at finish [B,kB,MB]: 26700639 26074.844 25.464 Starting time : 2006/01/02 21:50:15.029 Closing time : 2006/01/02 21:50:35.218 Total CPU time [seconds] : 20.18 I unerstand distance table and know which region of my alignment this points to but I do not understand the missing atoms or the out of range error, as I have 4142 atoms and no missing atoms. Could anyone help with where I am going wrong?! And also tell me if my procedure is correct, should the alignment automatically include the 3 BLK/water residues in the resdiue number if it is in the pdb file? and if i want the ligands in the output model is it correct to state /..w* at the end of this sequence in the alignment too? Also in the alignment is it correct for the seqeunce line to read: sequence:CcHh1: : : : :::-1.00:-1.00 ? Any feedback would be greatly appreciated. Thanks Jo ALIGNMENT >P1;Bamu structureX:Bamu: 1 : : 517 : :undefined:undefined:-1.00:-1.00 GTHEEEQYLFAVNNTKAEYPRDKTIHQLFEEQVSKRPNNVAIVCENEQLTYHELNVKANQLARIFIEKGIGKDTL VGIMMEKSIDLFIGILAVLKAGGAYVPIDIEYPKERIQYILDDSQARMLLTQKHLVHLIHNIQFNGQVEIFEEDT IKIREGTNLHVPSKSTDLAYVIYTSGTTGNPKGTMLEHKGISNLKVFFEN-SLNVTEKDRIGQFASISFDASVWE MFMALLTGASLYIILKDTINDFVKFEQYINQKEITVITLPPTYVVHL-----DPERILSIQTLITAGSATSPSLV NKWKE--KVTYINAYGPTETTICATTWVATKETIGHSVPIGAPIQNTQIYIVDENLQLKSVGEAGELCIGGEGLA RGYWKRPELTSQKFVDNPFVP-GEKLYKTGDQARWLSDGNIEYLGRIDNQVKIRGHRVELEEVESILLKHMYISE TAVSVHKDHQEQPYLCAYFVSE---KHIPLEQLRQFSSEELPTYMIPSYFIQLDKMPLTSNGKIDRKQLPEPDLT F/..w* >P1;CcHh1 sequence:CcHh1: : : : :::-1.00:-1.00 GPAERLELLAGWNDTDADRAR-PGLVEMFREQARARPDAVAVVDERRTLTYRQVEKLSAQLAHQLLARGLAVEDV VGISLERSAEMVIGLLAVLRAGGAFVPLDPHWPAERRAVVIEDAGVVVQLDASGEPAPG---EPEAVAVDLGDWR FGDRPTEDTGVTVPGDALAYVIFTSGSTGRPKGAMIRHEAISERLLWQIHEILGFGHDDASLFKAPLSFDISINE IFLPLVSGGRLVVLRPGGERDPHHLLSVIDEQRVTFTYLVSSMLDVLLEMAGDSGRLDSLRHVWCGGEVLTPELY ERFRTRLDIPLYHGYGPAETTIGVSHVVYRGAAERLSTSIGRANPNTRLYVLDDELRPVPVGVGGELYAGGLLLG RGYVNAPGLTASRFVANPFADDGSRLYRTGDLARFAPDGSLDFLGRADNQVKIRGMRLEIEDVEVGLAEHPRVRH TCVVARKNTAGGTYLVGYVIPAAGHEDLRADEVKAWAGEHMVEYMVPTHVVVMTEFPLTANGKLDRNALPEPVIP A/..w* MODLE.TOP INCLUDE SET ALNFILE = 'CcHh1-Bamu.ali' SET KNOWNS = 'Bamu' SET SEQUENCE = 'CcHh1' SET STARTING_MODEL = 1 SET ENDING_MODEL = 1 SET HETATM_IO = ON, WATER_IO = ON CALL ROUTINE = 'model'

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