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Re: [phenixbb] Rwork > Rfree
by Ed Pozharski 20 Nov '14

20 Nov '14

Generate a different test set and repeat refinement. Do it twenty times. If R turns out to be systematically higher than Rfree even in few resolution shells, you have an interesting case worth looking at. If not, it's just noise. Sent on a Sprint Samsung Galaxy S® III <div>-------- Original message --------</div><div>From: "Singh, Bishal" <b.singh(a)dkfz-heidelberg.de> </div><div>Date:11/20/2014 9:52 AM (GMT-05:00) </div><div>To: phenixbb(a)phenix-online.org </div><div>Subject: [phenixbb] Rwork > Rfree </div><div> </div>Hello everybody, While depositing a structure in PDB, I observed that Rwork was more than Rfree in the highest resolution shell and in a few more shells as well. I shall be delighted if somebody can suggest me the reasons and also what should I do now. With regards, Bishal PhD student, DKFZ, University of Heidelberg, Germany _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org http://phenix-online.org/mailman/listinfo/phenixbb

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GeoStd: was definition of glycosylation in phenix.refine
by Schubert, Carsten [JRDUS] 18 Nov '14

18 Nov '14

Hi Nigel, could you please comment on any plans to harmonize the use of these restraints for other refinement programs as well. Phenix originally appears to have implemented the restraint dictionaries from CCP4. Right now there seems to be some kind of parallel development going on with the dictionaries being in sync to some degree. Is there a plan to move to a unified restraints distribution at some point? Thanks Carsten From: phenixbb-bounces(a)phenix-online.org [mailto:[email protected]] On Behalf Of Nigel Moriarty Sent: Tuesday, November 18, 2014 12:38 PM Cc: PHENIX user mailing list Subject: Re: [phenixbb] definition of glycosylation in phenix.refine @Laurent Engin advice is sound but you are having trouble free feel to send me the inputs directly. NB. Any files sent to me will be held in strictest confidence. @Engin I think you will find what you are looking for in the GeoStd. As restraints are improved, they are moved from $PHENIX/chem_data/mon_lib to $PHENIX/chem_data/geostd (which is hosted on SourceForge for anybody's use). The directory structure is the same as the Monomer Library with a geostd_list.cif file corresponding to mon_lib_list.cif which specifies the links. Cheers Nigel --- Nigel W. Moriarty Building 64R0246B, Physical Biosciences Division Lawrence Berkeley National Laboratory Berkeley, CA 94720-8235 Phone : 510-486-5709 Email : NWMoriarty(a)LBL.gov<mailto:[email protected]> Fax : 510-486-5909 Web : CCI.LBL.gov<http://CCI.LBL.gov> On Tue, Nov 18, 2014 at 8:25 AM, Engin Özkan <eozkan(a)uchicago.edu<mailto:[email protected]>> wrote: Hi Laurent, If I am reading the error message right, you did not delete the hydrogens that get removed upon bond formation. That's what it is complaining about. I always thought Phenix probably removed these atoms (the hydroxyl and the hydrogen) automatically when you defined the link, but never tested if it really does it. I also usually remove all hydrogens right away when sugar residues are inserted. Finally, I will recommend that you use automatic linking in the phenix.refine GUI. It works great (I am on the latest nightly, dev-1839; you may need to upgrade). No need to define pesky links anymore. Thank you developers! At the risk of hijacking the thread, I could not find in my installation the cif files for N-linked sugars. Have they been moved elsewhere? Hope this helps, Engin On 11/18/14 10:02 AM, Laurent Maveyraud wrote: Hi, I am currently refining a structure of a glycosylated protein, but I fail to define the appropriate links... The glycosylatin is Asn638-NAG1061-(beta1-4)-NAG1062-(beta1-4)-BMA1063-(ALPHA1-3)-MAN1064 This is what I add in the eff file : apply_cif_link { data_link = NAG-ASN residue_selection_1 = chain D and resname NAG and resid 1061 residue_selection_2 = chain A and resname ASN and resid 638 } apply_cif_link { data_link = BETA1-4 residue_selection_1 = chain D and resname NAG and resid 1062 residue_selection_2 = chain D and resname NAG and resid 1061 } apply_cif_link { data_link = BETA1-4 residue_selection_1 = chain D and resname BMA and resid 1063 residue_selection_2 = chain D and resname NAG and resid 1062 } apply_cif_link { data_link = ALPHA1-3 residue_selection_1 = chain D and resname MAN and resid 1064 residue_selection_2 = chain D and resname BMA and resid 1063 } this is correctly read in refine: apply_cif_link: data_link: NAG-ASN mod_id_1: DEL-O1 mod_id_2: DEL-HD22 residue_selection_1: chain D and resname NAG and resid 1061 residue_selection_2: chain A and resname ASN and resid 638 apply_cif_link: data_link: BETA1-4 mod_id_1: DEL-HO4 mod_id_2: DEL-O1 residue_selection_1: chain D and resname NAG and resid 1062 residue_selection_2: chain D and resname NAG and resid 1061 apply_cif_link: data_link: BETA1-4 mod_id_1: DEL-HO4 mod_id_2: DEL-O1 residue_selection_1: chain D and resname BMA and resid 1063 residue_selection_2: chain D and resname NAG and resid 1062 apply_cif_link: data_link: ALPHA1-3 mod_id_1: DEL-HO3 mod_id_2: DEL-O1 residue_selection_1: chain D and resname MAN and resid 1064 residue_selection_2: chain D and resname BMA and resid 1063 but generates an error : Number of atoms with unknown nonbonded energy type symbols: 5 "HETATM10951 HO4 NAG D1052 .*. H " "HETATM10973 HO4 BMA D1053 .*. H " "HETATM11028 HO4 NAG D1062 .*. H " "HETATM11050 HO4 BMA D1063 .*. H " "HETATM11070 HO3 MAN D1064 .*. H " and the job stops. Any ideas on what if wrong ? thanks for your help Laurent _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org<mailto:[email protected]> http://phenix-online.org/mailman/listinfo/phenixbb

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definition of glycosylation in phenix.refine
by Laurent Maveyraud 18 Nov '14

18 Nov '14

Hi, I am currently refining a structure of a glycosylated protein, but I fail to define the appropriate links... The glycosylatin is Asn638-NAG1061-(beta1-4)-NAG1062-(beta1-4)-BMA1063-(ALPHA1-3)-MAN1064 This is what I add in the eff file : apply_cif_link { data_link = NAG-ASN residue_selection_1 = chain D and resname NAG and resid 1061 residue_selection_2 = chain A and resname ASN and resid 638 } apply_cif_link { data_link = BETA1-4 residue_selection_1 = chain D and resname NAG and resid 1062 residue_selection_2 = chain D and resname NAG and resid 1061 } apply_cif_link { data_link = BETA1-4 residue_selection_1 = chain D and resname BMA and resid 1063 residue_selection_2 = chain D and resname NAG and resid 1062 } apply_cif_link { data_link = ALPHA1-3 residue_selection_1 = chain D and resname MAN and resid 1064 residue_selection_2 = chain D and resname BMA and resid 1063 } this is correctly read in refine: apply_cif_link: data_link: NAG-ASN mod_id_1: DEL-O1 mod_id_2: DEL-HD22 residue_selection_1: chain D and resname NAG and resid 1061 residue_selection_2: chain A and resname ASN and resid 638 apply_cif_link: data_link: BETA1-4 mod_id_1: DEL-HO4 mod_id_2: DEL-O1 residue_selection_1: chain D and resname NAG and resid 1062 residue_selection_2: chain D and resname NAG and resid 1061 apply_cif_link: data_link: BETA1-4 mod_id_1: DEL-HO4 mod_id_2: DEL-O1 residue_selection_1: chain D and resname BMA and resid 1063 residue_selection_2: chain D and resname NAG and resid 1062 apply_cif_link: data_link: ALPHA1-3 mod_id_1: DEL-HO3 mod_id_2: DEL-O1 residue_selection_1: chain D and resname MAN and resid 1064 residue_selection_2: chain D and resname BMA and resid 1063 but generates an error : Number of atoms with unknown nonbonded energy type symbols: 5 "HETATM10951 HO4 NAG D1052 .*. H " "HETATM10973 HO4 BMA D1053 .*. H " "HETATM11028 HO4 NAG D1062 .*. H " "HETATM11050 HO4 BMA D1063 .*. H " "HETATM11070 HO3 MAN D1064 .*. H " and the job stops. Any ideas on what if wrong ? thanks for your help Laurent -- ---------------------------------------------------------- Laurent Maveyraud laurent.maveyraud AT ipbs DOT fr P I C T --- Plateforme Intégrée de Criblage de Toulouse Université Paul Sabatier / CNRS / I.P.B.S. UMR 5089 Département Biologie Structurale et Biophysique http://cribligand.ipbs.fr http://www.ipbs.fr 205 route de Narbonne 31077 TOULOUSE Cedex FRANCE Tél: +33 (0)561 175 435 Fax : +33 (0)561 175 994 ----------------------------------------------------------

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mrage with nucleic acids?
by Paul Paukstelis 17 Nov '14

17 Nov '14

I'm trying to run mrage with nucleic acid models. With phaser, it is possible to specify the molecule type, but this option doesn't exist in mrage (from what I can see). Providing a sequence file automatically assumes protein one letter codes. Any suggestion on how to clearly define a sequence file as a nucleic acid? Thanks, --paul

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Filtering CCP4 maps to lower resolution
by Barad, Ben 16 Nov '14

16 Nov '14

Hi All, I am currently trying to filter a CCP4-formatted real space map from the EMDB, which is at 3.2 Å resolution, to different resolutions from 3.2 to 5 Å, and I've been having a bit of a difficult time getting it to work. My first thought was to do an fft to get structure factors, then regenerate the map from those structure factors with a resolution cutoff. To do this, I used the tools "phenix.map_to_structure_factors" and "phenix.mtz2map" to generate a structure factor file, then convert it back into a map. When I do this, I end up with a very noisy map that is not usable for what I would like to do. Is there a better way to accomplish this in phenix (or another tool)? I've also tried some low-pass filtering with proc3d, without much success... Thanks, Ben Barad Fraser Lab @ UCSF

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Re: [phenixbb] simulated annealing omit map
by Pavel Afonine 15 Nov '14

15 Nov '14

Hello Bishal, as discussed before, computing ligand-omit map is tricky. Quoting my previous email: """ There are two commonly used options for omitting the ligand in order to demonstrate its presence/absence in Fo-Fc OMIT map: 1) Physically remove the ligand from PDB file. Then do some refinement and calculate Fo-Fc map. 2) Keep ligand in the file, set its occupancy to zero. Then, again, do some refinement and calculate Fo-Fc map. In this case you may want to ask refinement program to not move the ligand or move it only a little. Now, here is why these two options are poor and will not give you what you want. In the first case the bulk-solvent mask will be set in the ligand region and therefore it will mask ligand density (bulk-solvent will be filled into the ligand region). Depending on the strength of ligand density it may be masked completely or deteriorated. If you follow the second option you will always get positive density in ligand area. This density may correspond to bulk-solvent, ligand or mixture of both. That is there will be no simple way to differentiate whether this density arises from the ligand or bulk-solvent. """ Dale Tronrud offered a great alternative option: """ An alternative you might want to consider is what I call the "discovery map". At some point in the refinement process there was a map that convinced YOU that this ligand was present. You should be the hardest person to be convinced so that map will be both an omit map (because the model had been refined without the ligand prior to this) and clear enough to satisfy the reader. """ Now, recently I improved composite OMIT map calculation in phenix.composite_omit_map tool, see picture that illustrates how it works: https://www.dropbox.com/s/dbp8e0348v5p78h/fig_6.png?dl=0 I think at present that's the best option to follow if for some reason you cannot follow Dale's suggestion. Pavel On 11/13/14 6:58 AM, Singh, Bishal wrote: > Hello everybody, > I am generating figures showing the electron density map around the > ligands. I deleted the ligands from final co-ordinate file and then > performed refinement with identical parameters as before while > keeping simulated annealing=true. I shall be thankful if someone could > suggest me whether I consider mFo-DFc map or 2mFo-DFc map or both. > Kindly also tell me the minimum acceptable contour level required for > showing difference map. All structures are at 1.4 - 2.0 angstrom > resolutions. > > Regards, > Bishal

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CB outliers
by George Devaniranjan 14 Nov '14

14 Nov '14

Hi, I refined a structure using phenix.refine. The R/R free decreased slightly from 13.46/15.87 to 13.0/15.6. But, molprobity said there were 3 CB outliers (starting from 0). I looked at those residues in COOT. I could NOT really see a different between the refined and initial model. In addition I looked at COOT validate-->density fit analysis I cannot see ANY difference in the peak heights (same numerical value too). I looked at RSCC (using phenix validate tools as well). I still can't see a difference in the plots. I am not sure how to get rid of the 3 CB outliers that have crept in since I can't pick them out as worse off in the refined model. I ask since in another case > 3 CB outliers, with the same results in picking them out in COOT/RSCC. I am wondering if there is another way I can see the deviation and fix it? Thank you, George

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Rosetta compile problem
by miyatake 13 Nov '14

13 Nov '14

Dear all, I tried to compile Rosetta for rosetta_refinement, on Mac OX10.10. I run " /rosetta_2014.22.56873_bundle/main/source] user% sudo scons.py But it was in failure, showing the following message: g++-4.2 -o build/external/debug/macos/14.0/64/x86/gcc/4.2/default/dbio/cppdb/atomic_counter.os -c -isystem external/boost_1_55_0/ -isystem external/include/ -isystem external/dbio/ -pipe -ffor-scope -Wall -Wextra -pedantic -Werror -Wno-long-long -Wno-strict-aliasing -m64 -march=nocona -mtune=generic -O0 -g -ggdb -ffloat-store -fPIC -DBOOST_ERROR_CODE_HEADER_ONLY -DBOOST_SYSTEM_NO_DEPRECATED -DCPPDB_EXPORTS -DCPPDB_LIBRARY_SUFFIX=\".dylib\" -DCPPDB_LIBRARY_PREFIX=\"lib\" -DCPPDB_DISABLE_SHARED_OBJECT_LOADING -DCPPDB_DISABLE_THREAD_SAFETY -DCPPDB_SOVERSION=\"0\" -DCPPDB_MAJOR=0 -DCPPDB_MINOR=3 -DCPPDB_PATCH=0 -DCPPDB_VERSION=\"0.3.0\" -DCPPDB_WITH_SQLITE3 -Ibuild/external/debug/macos/14.0/64/x86/gcc/4.2/default/dbio/sqlite3 -Iexternal/dbio/sqlite3 -Iexternal/boost_1_55_0 -Iexternal/dbio -I/usr/include -I/usr/local/include external/dbio/cppdb/atomic_counter.cpp sh: g++-4.2: command not found scons: *** [build/external/debug/macos/14.0/64/x86/gcc/4.2/default/dbio/cppdb/atomic_counter.os] Error 127 scons: building terminated because of errors. How can I resolve that? Best regards, H. Miyatake

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simulated annealing omit map
by Singh, Bishal 13 Nov '14

13 Nov '14

Hello everybody, I am generating figures showing the electron density map around the ligands. I deleted the ligands from final co-ordinate file and then performed refinement with identical parameters as before while keeping simulated annealing=true. I shall be thankful if someone could suggest me whether I consider mFo-DFc map or 2mFo-DFc map or both. Kindly also tell me the minimum acceptable contour level required for showing difference map. All structures are at 1.4 - 2.0 angstrom resolutions. Regards, Bishal

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Settings-Directories of MR-Rosetta wizard is altered when phenix-GUI is restarted?
by Martin Moche 13 Nov '14

13 Nov '14

Dear colleagues, When restarting phenix followed by opening the "MR-Rosetta(beta)" the Settings-Directories need to be reset each time? How can I set Rosetta binary directory name: Rosetta binary name: Rosetta script directory name: Rosetta database directory name: in Settings-Directories of "MR-Rosetta(beta)" wizard (or elsewhere) so that they do not change each time I restart the phenix-GUI? These settings are not changed: Base output folder: Rosetta path: Likely "Rosetta path" does not change between phenix-GUI restarts since PHENIX_ROSETTA_PATH is set in the shell as: echo $PHENIX_ROSETTA_PATH /software/apps/rosetta/2014.35/gcc01/rosetta_2014.35.57232_bundle The "Path to Rosetta" in "Phenix preferences - Wizards" is currently left blank since $PHENIX_ROSETTA_PATH is set in the shell. I have by accident started a few jobs with mixed (new and old) settings and phenix-GUI starts without error messages and all processors are running 100% CPU, however likely in some infinite loop since no work is performed beyond "place model"... Best regards, Martin Martin Moche, Ph.D. Head of Protein Crystallography Karolinska Institutet MBB/PSF Scheeles väg 2 171 77 Stockholm Sweden phone: +46-8-524 868 43 mobile: +46-73-322 93 27 fax: +46-8-524 868 68 email: Martin.Moche(a)ki.se

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