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2nd announcement: 2013 workshop on Neutron Scattering Applications in Structural Biology
by Meilleur, Flora 02 Apr '13

02 Apr '13

4th Workshop on Neutron Scattering Applications in Structural Biology Oak Ridge, TN (USA). June 24 – June 28, 2013 Application deadline: April 22, 2013 No registration fee. Travel and accommodation grants available for participants from U.S. institutions. Detailed information available at: http://neutrons.ornl.gov/conf/gcnb2013/ Best Regards, Flora Meilleur, Ph. D Assistant Professor, Molecular and Structural Biochemistry North Carolina State University Neutron Sciences Directorate Oak Ridge National Laboratory Phone: 865-242-5747

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Fitting ligand using coot
by bsubedi＠btk.fi 31 Mar '13

31 Mar '13

Hi all, I tried to fit the ligand using the coot -> calculate-> other modeling tools-> Find ligands. I have imported cif to coot and also opened the pdb file of my ligand. However, coot do not identify the ligand and asks to adjust r.m.s.d. I can see the density for my ligand. Could anyone suggest me what could be the problem or if I am missing anything here. Best regards. Bishwa

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Re: [phenixbb] question about FFT map coefficients
by Michael Thompson 29 Mar '13

29 Mar '13

Hi Nat, The FFT didn't work out, but I'm not sure if it's due to negative Fs. When I set up the FFT map coefficients utility with my mtz file, after picking my input I get a warning that "The reflections file does not appear to contain any suitable map coefficients." Then if I try to run it anyway I get the error message "There are no map coefficients in this MTZ file." An mtz dump confirms that I have columns FP and SIGFP. Additional info: these Fs were generated using a python script that I wrote, and there are not meaningful values for SIGFP. My script simply wrote out 0.000 to the SIGFP column for each reflection essentially as a place holder. I thought that maybe 0 was not a valid entry for SIGFP so I changed it to 0.001 so there would be some value there, but I get all the same errors. Could this be the problem? Any suggestions? Thanks for the help. Sorry to bug you. Mike ----- Original Message ----- From: "Nathaniel Echols" <nechols(a)lbl.gov> To: "PHENIX user mailing list" <phenixbb(a)phenix-online.org> Sent: Thursday, March 28, 2013 3:42:31 PM GMT -08:00 US/Canada Pacific Subject: Re: [phenixbb] question about FFT map coefficients On Thu, Mar 28, 2013 at 3:28 PM, Michael Thompson <miket(a)chem.ucla.edu> wrote: > I have an mtz file with map coefficients that are essentially Fo-Fo for two data sets. I would like to generate a map using these coefficients, but because the subtraction of Fs has already been done, there are some values of FP that are negative. Is "FFT map coefficients" able to handle negative values for FP? My guess would be that the answer is yes, because internally the amplitudes and phases for map calculation are combined to form a single array of complex numbers, and this effectively converts the amplitudes to be all-positive. Also, if this were not the case, the isomorphous or anomalous difference maps would never work. But I haven't actually tried what you're wanting to do with real data - if the result isn't what you expect, let us know. -Nat _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org http://phenix-online.org/mailman/listinfo/phenixbb -- Michael C. Thompson Graduate Student Biochemistry & Molecular Biology Division Department of Chemistry & Biochemistry University of California, Los Angeles miket(a)chem.ucla.edu

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question about FFT map coefficients
by Michael Thompson 28 Mar '13

28 Mar '13

Hello All, A quick question about the "FFT map coefficients" utility in the GUI: I have an mtz file with map coefficients that are essentially Fo-Fo for two data sets. I would like to generate a map using these coefficients, but because the subtraction of Fs has already been done, there are some values of FP that are negative. Is "FFT map coefficients" able to handle negative values for FP? If not, I can always change the sign of negative Fs and add 180 to the corresponding model phases, but I was hoping I could avoid this. Thanks in advance for the help! Mike P.S. I do know there is an Fo-Fo map option in phenix, but the origin of my map coefficients is more complicated than simply Fo-Fo, so I cannot use this option. -- Michael C. Thompson Graduate Student Biochemistry & Molecular Biology Division Department of Chemistry & Biochemistry University of California, Los Angeles miket(a)chem.ucla.edu

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suggested usage of qsub?
by Scott Classen 28 Mar '13

28 Mar '13

Hi all, We have a HPC cluster configured with TORQUE/MAUI. I see in the Phenix GUI that there is some sort of ability to queue jobs to run via qsub. I can't find any documentation on how to set this up. If I'm working at my office computer and I want to submit to a remote server cluster for refinement, is this possible? Or is it assumed that qsub is available on the local machine? Thanks, Scott

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fwd: Deadline for the CCP4 Summer School application is approaching
by Nathaniel Echols 26 Mar '13

26 Mar '13

> ---------- Forwarded message ---------- > From: "Ruslan Sanishvili (Nukri)" <nukri(a)aps.anl.gov> > To: phenixbb(a)phenix-online.org > Date: Tue, 26 Mar 2013 11:52:44 -0500 (CDT) > Subject: Deadline for the CCP4 Summer School application is approaching > Dear Colleagues, > > The deadline for applications for the 6th annual CCP4 Summer School “From data collection to structure refinement and beyond” is only ten days away. Please submit your applications before April 5, 2013. The school will take place from June 18 to June 26, 2013 at the Advanced Photon Source (APS) near Chicago. > > There is no registration fee for the school. The students will be responsible for their own travel and lodging expenses. These and other details (the program, the list of speakers, the application process, accommodations, site access, contacts etc) can be found at the workshop website at http://www.ccp4.ac.uk/schools/APS-2013/index.php > > The school will include data collection, processing, structure solution, model building, refinement, validation, automation of many steps etc. Participants are encouraged to bring their own crystals, raw data or processed data for hands-on problem solving under the guidance of software developers and other experts. > We hope to see you in June > > Garib, Ronan and Nukri > > > > Ruslan Sanishvili (Nukri) > Macromolecular Crystallographer > GM/CA@APS > X-ray Science Division, ANL > 9700 S. Cass Ave. > Argonne, IL 60439 > > Tel: (630)252-0665 > Fax: (630)252-0667 > rsanishvili(a)anl.gov > >

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low-res. real-space refinement
by "Weiergräber, Oliver H." 26 Mar '13

26 Mar '13

Hello, when refining a protein structure against a low-resolution dataset (5 A), I wanted to also attempt real-space refinement. However, in the log file I find: |-real space refinement (target=simple)---------------------------------------| rejected... | | | --------------------------------------------------------------------------- | Changing the target to, e.g., diff_map does not change anything; the message will still refer to the "simple" target. I guess this is due to an internal resolution limit, but that's only speculation since it seems to be undocumented. Indeed, there is probably not much to be gained by real-space refinement at this resolution (at least not in terms of side-chain conformations), but I wanted to give it a try anyway. Any comments on this issue? Regards, Oliver ================================================ PD Dr. Oliver H. Weiergräber Institute of Complex Systems ICS-6: Structural Biochemistry Tel.: +49 2461 61-2028 Fax: +49 2461 61-9540 ================================================ ------------------------------------------------------------------------------------------------ ------------------------------------------------------------------------------------------------ Forschungszentrum Juelich GmbH 52425 Juelich Sitz der Gesellschaft: Juelich Eingetragen im Handelsregister des Amtsgerichts Dueren Nr. HR B 3498 Vorsitzender des Aufsichtsrats: MinDir Dr. Karl Eugen Huthmacher Geschaeftsfuehrung: Prof. Dr. Achim Bachem (Vorsitzender), Karsten Beneke (stellv. Vorsitzender), Prof. Dr.-Ing. Harald Bolt, Prof. Dr. Sebastian M. Schmidt ------------------------------------------------------------------------------------------------ ------------------------------------------------------------------------------------------------

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Drag drop files
by Colin Levy 25 Mar '13

25 Mar '13

Hi all, I am running 1.8.2-1309 under OSX 10.7.5 but I lost the ability to drag and drop files between various phenix windows. Anyone else experiencing this or is it something specific to my setup? Thanks, Colin Manchester Protein Structure Facility MIB 1.025 Tel 0161 306 5185 c.levy(a)manchester.ac.uk<mailto:[email protected]>

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Refinement of B-factors at low resolution
by Florian Brückner 25 Mar '13

25 Mar '13

Dear all I am refining a structure at 3.2 A with phenix.refine using automatic optimization of target weights, solved by molecular replacement with a 2.1A structure of the same crystal form. Rwork=26%, Rfree=28%. I use individual atomic B-factor refinement. The obtained distribution of B-factors after refinement looks reasonable and quite similar to the distribution in the higher resolution model, which I suppose would be expected, since it is the same crystal form. However, the average B-factor does not change substantially, i.e. if I start with the B-factors from the high resolution model (average B-factor around 32 A**2) after 6 macrocycles the average B-factor is still around 32 A**2. If I set all B-factors to the Wilson B-factor determined by phenix.refine (66 A**2), after 6 macrocycles the distribution of B-factors looks again very similar to the high resolution MR model, but the average B-factor is still around 65 A**2. What should I make of this? Is this an expected behavior at this resolution, i.e. the fact that the absolute values of B-factors cannot be determined during refinement, although a reasonable distribution can be obtained as judged by plotting average B-factors vs. residue#? Since the average B-factor does not change much during refinement, which average B-factor should I start refinement with to end up with reasonable absolute values? I have manually determined the Wilson B-factor from the linear part of the Wilson plot (4.5-3.2 A) to be 97 A**2. This seems high to me. Would that be expected for the given resolution, given that the 2.1A structure of the same crystal form has 32 A**2? I should add, that there is not a substantial effect of the average refined B-factor on Rfree. Thanks for any comments or suggestions. Best regards Florian ------------------------------------------------------------- Dr. Florian Brückner Laboratory of Biomolecular Research (LBR) OFLG/102 Paul Scherrer Institut CH-5232 Villigen PSI Switzerland Tel.: +41-(0)56-310-2332 Email: florian.brueckner(a)psi.ch

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molecular replacemnt problem
by Appu kumar 24 Mar '13

24 Mar '13

Dear members, I am doing a molecular replacement of a transcription factor whose ligand binding structure(24000 Da) is available in PDB but not for the DNA binding(13000 Da). When i am searching for the two copies from ligand binding domain as a template model, i am getting very good solution but i am not getting any density for the DNA binding domain to build up in density. The space gorup is P 1 21 1 (4) and unit cell parameters are Unit Cell: 57.43 69.36 105.99 90.00 90.00 90.00. Please guide me how to get the complete model structure. Table below show the matthews statistics For estimated molecular weight 37000. Nmol/asym Matthews Coeff %solvent P(2.20) P(tot) _____________________________________________________________ 1 5.71 78.46 0.00 0.01 2 2.85 56.91 0.62 0.70 3 1.90 35.37 0.37 0.29 4 1.43 13.82 0.00 0.00 _____________________________________________________________ The phaser molecular replacement gives the following table. istogram of relative frequencies of VM values ---------------------------------------------- Frequency of most common VM value normalized to 1 VM values plotted in increments of 1/VM (0.02) <--- relative frequency ---> 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 | | | | | | | | | | | 10.00 - 8.33 - 7.14 - 6.25 - 5.56 - 5.00 - 4.55 - 4.17 - 3.85 -- 3.57 --- 3.33 ------ 3.12 ---------- 2.94 **************** (COMPOSITION*1) 2.78 ----------------------- 2.63 -------------------------------- 2.50 ----------------------------------------- 2.38 ------------------------------------------------ 2.27 -------------------------------------------------- 2.17 ----------------------------------------------- 2.08 -------------------------------------- 2.00 -------------------------- 1.92 --------------- 1.85 ------- 1.79 --- 1.72 - 1.67 - 1.61 - 1.56 - 1.52 - 1.47 * (COMPOSITION*2) 1.43 - 1.39 - 1.35 - 1.32 - 1.28 - 1.25 - TABLE : Cell Content Analysis: SCATTER :N*Composition vs Probability:0|3x0|1:1,2: $$ N*Composition Probability $$ loggraph $$ 1 0.306066 2 0.00141804 $$ Most probable VM for resolution = 2.27817 Most probable MW of protein in asu for resolution = 92664.2 I have initially indexed the data in C2221 but Rfree was not decreasing so i reindexed the data in data in P121 space group keeping the Rfree flag of C2221. While analysing the symmetry mates , i found large space but no density. structure of Ligand binding domain is almost identical with 90% identity in sequence. I am stuck with this problem and don't know how to process further. I run the phenix.xtriage to evaluate the twining but it suggest no twining. When i reindex from C2221 to P21, the completeness of data reduced from 95 % to 35% whereas the map is very good and Rwork and Rfree are 26/31 for 2.2 resolution. I do not understand why the completeness of data reduced so much on reindexing. please Can anyone explain this phenomenon. Please give me your valuable suggestion. I will appreciate your effort. Thank you

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one or more errant TER cards
by jp d 20 Mar '13

20 Mar '13

hi, i am trying to refine using a reference model and keep getting this error *** Adding Reference Model Restraints *** Sorry: chain ' ' in start.pdb contains one or more errant TER cards. Please remove and try again i've tried various things including removing all the TER lines my models (reference and working) has many chains identified by segid's any suggestions ? thanks jpd

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install on ubuntu 12.04?
by Alastair Fyfe 19 Mar '13

19 Mar '13

Hi, I've installed previous phenix builds on ubuntu without problem, but with both 1.8.1-1168 and dev-1269 installers on 12.04, the command "./install --prefix=/home/afyfe" ends with "Error configuring: see /home/afyfe/phenix-installer-dev-1269/build-binary/intel-linux-2.6/svr/log/binary.log for details" and the tail of the binary.log file suggests: -- Creating files in build directory: "/home/afyfe/phenix-dev-1269/build/intel-linux-2.6" dispatcher_include_template.sh dispatcher_head.sh Please re-configure from scratch your cctbx_build:cd cctbx_build; /your/path/to/python ../cctbx_project/libtbx/configure.py [options] [modules] --- I don't see a cctbx_build directory under either the installer or destination directories. Any clues/suggestions? many thanks, Alastair

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International Conference on Structural Genomics - Structural Life Science, Sapporo, Japan, July 29-Aug 1, 2013
by Terwilliger, Thomas C 18 Mar '13

18 Mar '13

Dear Colleagues: On behalf of the organizing committee of the International Conference on Structural Genomics 2013 – Structural Life Science – (ICSG2013-SLS), we cordially welcome you to the conference, to be held in Sapporo, Hokkaido, Japan, July 29th – August 1st, 2013. ICSG2013-SLS is intended to provide an overview for the most recent developments in Structural Genomics and its impact on research in biology, medicine and disease, and to foster international collaboration among researchers. You can see all the details of the conference at: http://www.c-linkage.co.jp/ICSG2013 . Registration for the conference is now open. The scientific topics covered in ICSG2013-SLS include the wider life science research fields with particular attention to drug discovery (small molecules and biopharmaceuticals), biotechnology and industrial issues while keeping strength in the high-throughput technologies and integration of hybrid methods. These technologies are now leading to the new field of “Structural Life Science”. ICSG2-13-SLS is partially supported by the Grant-in-Aid for Scientific Research on Innovative Areas; “Structural Cell Biology”, “Intrinsically Disordered Protein” and “Transient Macromolecular Complexes”, from Ministry of Education, Culture, Sports, Science and Technology (MEXT) . In order to widen the opportunity to young and enthusiastic fellows to study more, we have organized several satellite workshops before ICSG2013- SLS. The topics will include “Small molecule screening”, “Automation of X-ray Structure Determination”, “Cell-free Protein Production”, “Automated NMR methods”, "Eukaryotic expression", “Interaction analyses" and "Bioinformatics”. We hope you also find the satellite workshops are informative and productive. The conference will be held in Keio Plaza Hotel Sapporo, in walking distance of Hokkaido University's main campus and Sapporo station. The summer in Sapporo is a great time to stay and enjoy the cool summer night of Japan. We are looking forward to welcoming you to Sapporo in the summer of 2013. Sincerely yours, Katsumi Maenaka, Ph.D. Chair, International Conference on Structural Genomics 2013 -Structural Life Science- (ICSG2013-SLS) Laboratory of Biomolecular Science and Center for Research and Education on Drug Discovery, Faculty of Pharmaceutical Sciences, Hokkaido University, Japan Soichi Wakatsuki Chair of Program Committee, ICSG2013-SLS Photon Science, SLAC National Accelerator Laboratory Department of Structural Biology School of Medicine Stanford University

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MR_rosetta and disulphides
by Morten Groftehauge 17 Mar '13

17 Mar '13

Hi guys, Phenix version 1323 Rosetta version 3.4 I try to run phenix.mr_rosetta rosetta_command="-MR::disulf 129:411 228:281 279-615" mr_rosetta_228.eff but I get the following error in rebuild.log ERROR: Option matching -MR:disulf not found in command line top-level context I lifted the rosetta_command directly from the Version Documentation. Any clues as to why this doesn't work? Sincerely, -- Morten K Grøftehauge, PhD Pohl Group Durham University

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Tutorial of low resolution data?
by 李翔 17 Mar '13

17 Mar '13

Hi everyone, I will work on some low resolution data but have very little experience. (Data are about 4 A to 8 A) Does anyone know where I could find a tutorial for low resolution data please? It is best to work with Phenix, other software is fine. MIR or SAD or MR method will be appreciated and it is very helpful if the tutorial can talk about some details of the data processing. Cheers, Frank

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Phenix DEN-twin-refinement
by Ethayathulla A.S 14 Mar '13

14 Mar '13

hi all I want to do DEN twin-refinement using phenix. For my data phenix.DEN works better than CNS DEN. I want to DEN with twinning. Is there script for adding twinning operator for DEN refinement in phenix phenix.den_refine model.pdb data.mtz den.reference_file=reference.pdb Thank you Abdul --

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Re: [phenixbb] problems installing latest phenix on mac osx lion
by Alejandro Buschiazzo 14 Mar '13

14 Mar '13

Thanks Nat! read your message a bit later (I think I need to change from the digest format, so I receive each message as it comes), looking at http://phenix-online.org/pipermail/phenixbb/2013-March/thread.html Actually, your suggestion worked all right. (and by sourcing the phenix_env.sh as you point out, it's actually the same as the command line -installer of course) Don't know why the errors appeared before, but in any case, now it's running fine. Thank you for your help Alejandro > Not sure what's going on here - I will check it out today. There's > one more option however that should be the safest: did you try the > graphical installer yet? The destination can't be changed, but you > can still use the command line if you want by doing this: > > source /Applications/PHENIX-1.8.2-1309/Contents/phenix-1.8.2-1309/phenix_env.sh > > (or the csh equivalent) > > But I will see if there's an explanation for these errors. > > -Nat On Tue, Mar 12, 2013 at 4:26 AM, Alejandro Buschiazzo <alebus at pasteur.edu.uy <http://phenix-online.org/mailman/listinfo/phenixbb>> wrote: >/ I 'm trying to install phenix 1.8.2-1309 on a Macbook Pro (os x 10.7.5). />/ />/ First tried the binary installer proposed (which is indicated for os x />/ 10.6.7) />/ It didn't work, ending with "fix_mac_python: caught error - exiting." />/ />/ I spotted in the bb that this could've happened because the downloaded />/ installer was somehow corrupted, tried downloading it again, but same />/ result. />/ />/ Tried then the source installer: everything OK - except for wxPython, giving />/ this error />/ />/ WXPYTHON: />/ Using wxOSX_Cocoa build of wxPython 2.9 alpha version />/ />/ building wxPython-src-2.9.4.1 />/ ERROR: installation of wxosx_cocoa failed />/ />/ ERROR: installation of wxpython failed />/ />/ ...now I can work with command lines, but the GUI won't work of course. />/ />/ Your help is appreciated />/ />/ Alejandro />/ />/ />/ -- />/ Alejandro Buschiazzo, PhD />/ Research Scientist />/ Unit of Protein Crystallography />/ Institut Pasteur de Montevideo />/ Mataojo 2020 />/ Montevideo 11400 />/ URUGUAY />/ Phone: +598 25220910 int. 120 />/ Fax: +598 25224185 />/ http://www.pasteur.edu.uy/pxf/ -- Alejandro Buschiazzo, PhD Research Scientist Unit of Protein Crystallography Institut Pasteur de Montevideo Mataojo 2020 Montevideo 11400 URUGUAY Phone: +598 25220910 int. 120 Fax: +598 25224185 http://www.pasteur.edu.uy/pxf

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Fwd: 4th Edition of the ISBC2013
by Nathaniel Echols 14 Mar '13

14 Mar '13

---------- Forwarded message ---------- From: Jose A. Gavira <gavirius(a)gmail.com> Date: Thu, Mar 14, 2013 at 11:20 AM Subject: 4th Edition of the ISBC2013 Dear Colleague, The Laboratory of Crystallographic Studies is pleased to announce the 4th International School on Biological Crystallization (ISBC2013), to be held in Granada (Spain) during May 26-31, 2013. ISBC2013 is intended for postgraduate/postdoctoral students and research scientists from industrial and academic backgrounds. The International School will provide five days of lectures, posters and practical demonstrations focused on the fundamentals of crystallization from solution and their applications to the field of crystallization of biological materials, including biological macromolecules, biominerals and biomimetic materials. One day will be fully devoted to case studies on the crystallization of membrane proteins, viruses and large macromolecular complexes. For more information, please visit http://www.isbcgranada.org/<http://www.linkedin.com/redirect?url=http%3A%2F%2Fwww%2Eisbcgranada%2Eorg%2…> . Best regards, ISBC2103 Organizing Committee --- Gavi http://isbcgranada.org/

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problems installing latest phenix on mac osx lion
by Alejandro Buschiazzo 12 Mar '13

12 Mar '13

Hello, I 'm trying to install phenix 1.8.2-1309 on a Macbook Pro (os x 10.7.5). First tried the binary installer proposed (which is indicated for os x 10.6.7) It didn't work, ending with "fix_mac_python: caught error - exiting." I spotted in the bb that this could've happened because the downloaded installer was somehow corrupted, tried downloading it again, but same result. Tried then the source installer: everything OK - except for wxPython, giving this error > WXPYTHON: > Using wxOSX_Cocoa build of wxPython 2.9 alpha version > > building wxPython-src-2.9.4.1 > ERROR: installation of wxosx_cocoa failed > > ERROR: installation of wxpython failed ...now I can work with command lines, but the GUI won't work of course. Your help is appreciated Alejandro -- Alejandro Buschiazzo, PhD Research Scientist Unit of Protein Crystallography Institut Pasteur de Montevideo Mataojo 2020 Montevideo 11400 URUGUAY Phone: +598 25220910 int. 120 Fax: +598 25224185 http://www.pasteur.edu.uy/pxf

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GUI display items
by Luca Pellegrini 10 Mar '13

10 Mar '13

Hello, Is there a way to change/select the column items displayed in the PHENIX home window, under the 'Job history' tab? At the moment, mine shows: ID/Program/Date/Job title or directory/R-free. I would like to add things like the Overall score from MolProbity etc. I looked under Preferences but couldn't find anything. Thanks, Luca Luca Pellegrini Department of Biochemistry University of Cambridge 80 Tennis Court Road Cambridge CB2 1GA - UK Email: lp212(a)cam.ac.uk Tel: 0044-1223-760469 Fax: 0044-1223-766002 Sanger building, room 3.59

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fwd: EMBO 2013 Practical Course - Exploiting Anomalous Scattering in Macromolecular Structure Determination ESRF-EMBL, Grenoble, France, 10 - 14 June 2013
by Nathaniel Echols 09 Mar '13

09 Mar '13

(Note that the registration deadline is tomorrow.) ---------- Forwarded message ---------- From: Daniele de Sanctis <desancti(a)esrf.fr> To: phenixbb(a)phenix-online.org Date: Sat, 9 Mar 2013 12:51:54 +0100 Subject: EMBO 2013 Practical Course - Exploiting Anomalous Scattering in Macromolecular Structure Determination ESRF-EMBL, Grenoble, France, 10 - 14 June 2013 DEADLINE Tomorrow COURSE ANNOUNCEMENT EMBO 2013 Practical Course - Exploiting Anomalous Scattering in Macromolecular Structure Determination ESRF-EMBL, Grenoble, France, 10 - 14 June 2013 The EMBO 2013 Practical Course on Exploiting Anomalous Scattering in Macromolecular Structure Determination will be hosted by the ESRF in Grenoble, France from 10 to 14, June 2013. The course aims to impart the theoretical and practical basis for the 3-dimensional structure determination of bio-macromolecules using Anomalous Dispersion techniques (SAD & MAD) to young scientists who intend to apply these methods in macromolecular crystallography. Through a series of lectures, software demonstrations, practicals on the ESRF beamlines and tutorials, participants will get insights into all aspects of the structure determination process including beamline instrumentation, data collection and processing, heavy atom substructure determination, phasing and model building. There will also be sessions focusing on automated structure solution procedures and newer methods. Additional sessions will include presentations of distinguished structures solved with these techniques. The number of participants is limited to 20 and the deadline for application is March 10th, 2013. Additional information, course programme and instructions to apply to the course can be found in the course webpages: http://events.embo.org/13-crystallography/ Confirmed invited speakers include: Andreas Bracher, Kay Diederichs, Rita Giordano, Adrian Goldman, Wolfgang Kabsch, Gordon Leonard, Airlie McCoy, Sean McSweeney, Christoph Müller-Dieckmann, Max Nanao, Alexander Popov, Harry Powell, Thomas Terwilliger, Andrew Thompson, Isabel Uson, Clemens Vonhrein -- ἀρετή --------------------------------------- Daniele de Sanctis, PhD Structural Biology Group ESRF, Grenoble, France Tel 33 (0)4 76 88 2869

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map woes from refine
by Machius, Mischa Christian 08 Mar '13

08 Mar '13

Y'all, Recently, I've run into an issue with the maps from refine. All during the refinement cycles, the autoupdated maps in the associated coot window look great as refinement progresses. However, the final maps look horrible; completely chopped up. It's not a matter of contouring levels; it's essentially unrecognizable, and phenix acknowledges that by stating very low correlation coefficients at the end of the log file. I have downloaded the latest builds in an attempt to address B-value issues that were recently discussed and that I am (still) struggling with. The map issue happens with all of them. Is it possible that there is some crossfire from different versions, or any other reason? Any help appreciated. Cheers, MM

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selection of different residue ranges for a NCS group
by Andres PALENCIA 07 Mar '13

07 Mar '13

Dear All, I am trying to select different regions of the same protein to be included in the same NCS group before refinement. This correspond to the core of a protein. Additionally, I would define other NCS groups for external domains. I have 4 protein/RNA complexes in the asu, P1. Here are the residue ranges for the NCS group corresponding to the core of the protein (invariant): Chain A and resseq: 1:156, 188-227, 413:568, 629:794 this should be the reference group, to match the same region in the other 3 monomers (C, E. G). Other NCS groups will include the RNA, as well as other external domains: Chain B and resseq: 1: 87 This is the RNA molecule, that should match the other 3 monomers (D, F, H) Chain A resseq: 157:187 this is a external domain, to match the other protein monomer (C, E, G) and so on.... for other external domain I have seen that there is the possibility to use user-defined NCS groups, by using: * ncs_groups.params * or modifying the GUI by editing the atom selection For the external domains I think is easy as the syntax is only one line, however for the core of the protein I do not manage to select properly the different residue ranges. Could anybody please help on that? what would be the easiest way to do that? Can I for example creat a ncs_groups.params file and add it to phenix.refine in the GUI? Thanks in advance, Andres

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map scaling: sigma or volume scaled
by Edward A. Berry 07 Mar '13

07 Mar '13

map scaling can be defined: sigma or volume scaled Maybe another dumb question, but what is volume scaling? For 2mFo-DFc and mFo-DFc maps. Thanks, Ed

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quick clashlist or molprobity in batch mode
by jan 06 Mar '13

06 Mar '13

Hi all, I want to analyze a whole bunch of structures with molprobity, and I am looking for a convenient way to do this. phenix.quick_clashlist seems to be a handy tool. However, when I use the following command line: "phenix.quick_clashlist model=mypdb.pdb" ... the output tells me: # run_clashlist # # Run reduce and molprobity # type phenix.run_clashlist: --help for help Parameters used: run_clashlist { model = "4iuj.pdb" build = True summary_file = None output_file = None temp_dir = None remove_temp_dir = None verbose = False raise_sorry = False debug = False dry_run = True } ************************************************ 'str' object has no attribute '_anchor' ************************************************ Any ideas what I need to do differently? thanks, Jan -- Jan Abendroth Emerald BioStructures Seattle / Bainbridge Island WA, USA home: Jan.Abendroth_at_gmail.com work: JAbendroth_at_embios.com http://www.emeraldbiostructures.com

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Using Phenix Refine for Hypothetical Models
by Sam Stampfer 06 Mar '13

06 Mar '13

Dear Phenix Community, Is it possible to use phenix.refine to do simulated annealing and energy minimization of a hypothetical model? I am currently using pyrosetta relax for minimization; however, my hypothetical model is based on a homologous protein whose structure is a crystallographic trimer. I would like to preserve my model's symmetry, which I know is possible in phenix.refine but not in pyrosetta (the symmetry controls are only supported in regular rosetta, as far as I know). Is there a way to have phenix ignore x-ray data and just do energy minimization functions? How would I go about doing that? Would I need a file containing fake/dummy x-ray data? Thanks for your help! -Sam Stampfer

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Fwd: FINAL REMINDER: ALS Call for General User Proposals - DEADLINE March 6 2013
by Banumathi Sankaran 05 Mar '13

05 Mar '13

Dear Users, I am forwarding the reminder mail from ALS to submit your proposal for the CC time and general user time at ALS. Please submit your proposals before time. Thanks Banu FINAL REMINDER: CALL FOR GENERAL USER PROPOSALS AT THE ALS The User Services Office is accepting General User Proposals from scientists who wish to conduct research at the ALS in the next cycle. PROPOSAL SUBMISSION DEADLINE cycle: 2013-2 August-December Deadline: March 6, 2013 NEW PROPOSALS Please follow the instructions for proposal submission at: http://www-als.lbl.gov/index.php/user-information/user-guide/58.html BEAM TIME REQUEST for an ACTIVE PROPOSAL Proposals remain active for two years (up to four cycles), but beam time is only allocated if a beamtime request is made each cycle. After two years the proposal expires and a new proposal must be submitted. If you have an eligible active proposal you will already have received an email from us which includes your login number and password. PUBLICATION REMINDER The proposal form now includes a section on publications from previous ALS work. Please make sure your publications are entered into our database: http://alsusweb.lbl.gov/4DCGI/WEB_GetForm/PublicationSearch.shtml/Initialize If you have questions, please feel free to contact me. Cheers, Clyde Lewis Proposal Coordinator ALS

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Memory and CPU usage
by Jon Agirre 05 Mar '13

05 Mar '13

Dear community, I'm planning on buying a new macbook for my first abroad postdoc venture, and I'm currently trying decide what amount of RAM & CPU to choose. My question is a little bit intricate: is there any rule of thumb to estimate the needed RAM versus the number of input atoms for any particular refinement type? I'm planning on tackling some big (18,000 residues/asu) refinements but I don't have any estimation as to how many GBs of RAM they might require. The maximum amount of RAM I can install is 16GB, but it is a non-standard configuration that might void Apple's warranty. About the CPU: how scalable are typical phenix processes? Would it be sensible to invest in a quad-core machine with HT? In this particular case, and since HT would present 8 logical cores, would I get any speed up from launching phenix tasks configured for 8 processors instead of 4? Thanks in advance! Jon

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Unusually low B factors with phenix 1.8, but not with phenix 1.7 (data set has ice rings)
by Oliv Eidam 05 Mar '13

05 Mar '13

Hi, I would like to continue the B-factor discussion started by Demetres D. Leonidas on Nov 2, 2012 on CCP4BB: http://www.mail-archive.com/[email protected]/msg29012.html I make the same observations like Demetres and Mingzhu Wang: unusually low B-factors when refining with Phenix 1.8-1069 and 1.8.1-1168, but not with Phenix 1.7.1-743. In numbers: the average B-factor is 10 A2 but it should be 20 A2 (based on Wilson B and similar structures). A few atoms (~20) have Bf = 0.00. The following is important: I refined dozens of similar structures without that problem and I observe this phenomenon only for a single data set. That specific data set is different because 1) I excluded regions with ice rings in XDS and 2) I limited the resolution to 2.28 Angstroem, although the crystal diffracted to ~1.8 Angstroem. While the preparation of the data set might be questionable in itself, I would prefer that this would not be the topic of this thread. My questions are: What can I do to refine my structure with Phenix 1.8.X using this truncated data set? What causes the difference in average B-factor values between Phenix 1.7 and 1.8 for this data set? Your advice is much appreciated. Oliv On 11/04/12 19:21, mingzhu wang wrote: > Pavel, > > 1.8.1-1168 also has this problem. > > I found this problem existed with some dataset, but not all dataset. > > 1.8.1 give better map, so I built model with 1.8.1 map and go back to > 1.7.3 before I deposit the structure to PDB. > > When I compared the B factor from 1.8.x and 1.7.3, I found > B(1.7.3)=B(1.8.x)+Boverall(1.8.x). Perhaps this is the problem. > > > > 2012/11/3, Pavel Afonine<pafonine(a)gmail.com>: >> Joao, >> >> I am using the latest version of phenix 1.8.1-1168 >> >> >> 1.8.1-1168 should not have that "problem". >> >> If you suspect there is still a problem, you can send me the data and model >> files off list, explain what exactly the problem is, and I will have a look >> right away. >> >> FYI: there is Phenix mailing list for Phenix-specific questions, where I >> can also explain in great details the behavior with the B-factors you >> observe. >> >> All the best, >> Pavel >> -- Oliv Eidam, Ph.D. Postdoctoral fellow University of California, San Francisco Dept. of Pharmaceutical Chemistry 1700 4th Street, Byers Hall North, Room 501 San Francisco, CA 94158 - Box 2550 Phone: 415-514-4253 Fax : 415-514-4260 Email:[email protected]

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fwd: Reminder: CCP4 summer school at APS, USA
by Nathaniel Echols 04 Mar '13

04 Mar '13

---------- Forwarded message ---------- From: "Ruslan Sanishvili (Nukri)" <nukri(a)aps.anl.gov> To: phenixbb(a)phenix-online.org Date: Mon, 4 Mar 2013 16:14:43 -0600 (CST) Subject: Reminder: CCP4 summer school at APS, USA Dear Colleagues, This is a reminder that the deadline for applications for the 6th annual CCP4 Summer School “From data collection to structure refinement and beyond” is April 5, 2013. The school will take place from June 18 through June 26, 2013 at the Advanced Photon Source (APS) near Chicago. There is no registration fee for the school. The students will be responsible for their own travel and lodging expenses. These and other details (the program, the list of speakers, the application process, accommodations, site access, contacts etc) can be found at the workshop website at http://www.ccp4.ac.uk/schools/APS-2013/index.php The school will include data collection, processing, structure solution, model building, refinement, validation, automation of many steps etc. Participants are encouraged to bring their own crystals, raw data or processed data for hands-on problem solving under the guidance of software developers and other experts. We hope to see you in June Garib, Ronan and Nukri Ruslan Sanishvili (Nukri) Macromolecular Crystallographer GM/CA@APS X-ray Science Division, ANL 9700 S. Cass Ave. Argonne, IL 60439 Tel: (630)252-0665 Fax: (630)252-0667 rsanishvili(a)anl.gov

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Bay Area and Beyond Phenix User Meeting (March 14th)
by Paul Adams 04 Mar '13

04 Mar '13

Dear Colleagues, On Thursday March 14th, a Bay Area and Beyond Phenix User Meeting will be held at Lawrence Berkeley Lab. During the workshop, the Phenix developers will give lectures on the latest developments and Phenix usage. Half of the user meeting will be in the form of informal hands on sessions, allowing participants to work with the developers. Participants are encouraged to bring their own (reduced) data to be used during hands on sessions, but tutorial data will also be available. We also recommend bringing a relatively recent laptop computer (any OS). Due to the highly interactive format of the meeting there will be space for approximately 20 participants. If I have missed any of your colleagues on this invitation please feel free to pass on the message. To apply, researchers should send a short summary email describing their level of expertise and motivation for attending the workshop to NEchols(a)lbl.gov and PDAdams(a)lbl.gov by Friday March 8th. Cheers, Paul -- Paul Adams Deputy Division Director, Physical Biosciences Division, Lawrence Berkeley Lab Division Deputy for Biosciences, Advanced Light Source, Lawrence Berkeley Lab Adjunct Professor, Department of Bioengineering, U.C. Berkeley Vice President for Technology, the Joint BioEnergy Institute Laboratory Research Manager, ENIGMA Science Focus Area Building 64, Room 248 Tel: 1-510-486-4225, Fax: 1-510-486-5909 http://cci.lbl.gov/paul Lawrence Berkeley Laboratory 1 Cyclotron Road BLDG 64R0121 Berkeley, CA 94720, USA. Executive Assistant: Louise Benvenue [ LBenvenue(a)lbl.gov ][ 1-510-495-2506 ] --

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phenix.refine multiple twin laws?
by Kip Guja 03 Mar '13

03 Mar '13

Hi All, Is the newest version of Phenix capable of refinement with multiple twin laws (eg, tetartohedral twinning)? If so, any advice on input parameters, etc. would be greatly appreciated. Best wishes, Kip

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Re: [phenixbb] Magnesium or calcium
by Daniel Slade 02 Mar '13

02 Mar '13

So the resolution is 1.6 and I remember the B factor for mg at 16. I haven't checked the coordinating residues. It coordinates acidic residues, a backbone carbonyl and one water. I'm now leaning towards calcium as the other calciums have bonds distances similar to this one, but not nearly as high an affinity. Thanks

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Magnesium or calcium
by Daniel Slade 02 Mar '13

02 Mar '13

I have several structures that has multiple known calcium binding sites, and I found a new site that seems to accept calcium or magnesium. I say this because if a magnesium is put in the structure, the bond lengths are ~ 2.1. But if you put a calcium in it, they go to ~2.6. All of these refinements were done without ReadySet, which I recently used with metal restraints and it seemed to change the bond length of calcium to ~2.4. I'm a bit confused as to what to do here as either metal seems to be right. Worth noting, this is a very strong metal binding site that can't be removed with 10 mM EDTA. Any help here on how to figure this out which ion this is would be great. Thanks in advance Dan

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Re: [phenixbb] Magnesium or calcium
by Ed. Pozharski 02 Mar '13

02 Mar '13

Magnesium has fewer electrons. So if you place it and it's actually calcium you should see extra density and the B-factor will be unreasonably lower than surrounding atoms. Distances are less reliable at intermediate resolutions as they are largely controlled by restraints. Maybe. Assuming that you have 100% occupancy. -------- Original message -------- From: Daniel Slade <DSlade(a)scripps.edu> Date: To: phenixbb(a)phenix-online.org Subject: [phenixbb] Magnesium or calcium I have several structures that has multiple known calcium binding sites, and I found a new site that seems to accept calcium or magnesium. I say this because if a magnesium is put in the structure, the bond lengths are ~ 2.1. But if you put a calcium in it, they go to ~2.6. All of these refinements were done without ReadySet, which I recently used with metal restraints and it seemed to change the bond length of calcium to ~2.4. I'm a bit confused as to what to do here as either metal seems to be right. Worth noting, this is a very strong metal binding site that can't be removed with 10 mM EDTA. Any help here on how to figure this out which ion this is would be great. Thanks in advance Dan _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org http://phenix-online.org/mailman/listinfo/phenixbb

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question about reflection file tools behavior
by Michael Thompson 01 Mar '13

01 Mar '13

Hello All, I have a question about the behavior of the reflection file tools within the PHENIX GUI: I am trying to simply copy a set of FreeRFlags from a higher resolution dataset to a lower resolution dataset. The low-res dataset contains 35264 reflections in the original input mtz. The high-res set contains 43731 reflections. When I use the GUI to write a new mtz using the low-res amplitudes and the high-res FreeRFlags, the result is an mtz with 46323 reflections. I am confused about why the output has so many reflections, even more than the high res dataset? Also, I noticed that while both input mtz files have a low-res cutoff of 20A, the output mtz has a low-res cutoff around 75A. Can anyone please explain what is going on here? I would like my output file to have the same reflections as the input, and no more. I have noticed that the CCP4 program FreeRFlag seems to behave in a similar way, so my guess is that what is happening is a common procedure, but I'm at a loss for an explanation of why this is the case. Also, if there's a better way to implement what I am trying to do I'd be grateful to hear it (perhaps generate entirely new flags for both datasets using the same random seed?). Thanks for the help!! Mike -- Michael C. Thompson Graduate Student Biochemistry & Molecular Biology Division Department of Chemistry & Biochemistry University of California, Los Angeles miket(a)chem.ucla.edu

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mr_rosetta fails after autobuild stage
by Huw Jenkins 01 Mar '13

01 Mar '13

Hi, I've been having some problems with mr_rosetta. All the regression tests run fine but when I run it on some real data it gets as far as the autobuilding stage and then fails with an error message. All I can find in the log files GROUP_OF_AUTOBUILD_1/RUN_FILE_1.log GROUP_OF_AUTOBUILD_1/RUN_FILE_2.log is ******************* ERROR ENDING *************** Failed to carry out autobuild ******************* ERROR ENDING *************** The autobuild jobs themselves seem to finish fine i.e the end of the log files in GROUP_OF_AUTOBUILD_1/RUN_1/AutoBuild_run_1_ and GROUP_OF_AUTOBUILD_1/RUN_2/AutoBuild_run_1_ has the tables of Rfactors per cycle and the list of citations with no indiction of any error. Is there anywhere else I should be looking for a more detailed error message? Thanks, Huw

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real_space_correlation
by Jianghai Zhu 28 Feb '13

28 Feb '13

Hi All, I was trying to use "phenix.real_space_correlation *.pdb resolve_composite_map.mtz" to get some real space cc for my ligand against the SA composite omit map. But I am not sure that the map used here was the SA composite omit map. Will phenix use the omit map I provided or will it just simply calculate a sigmaA weighted 2Fo-Fc map based on the pdb? Thanks. --Jianghai

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mr_rosetta: two proteins, one chain placed, build second chain
by Tobias Beck 28 Feb '13

28 Feb '13

Dear phenixbb, I would like to use mr_rosetta to build a second protein chain. For the first protein, I have a search model with known structure. The search model can be placed with Phaser (TFZ 10). For the second protein, the structure of a homologue has been determined. The second protein accounts for about 2/3 of the residues in the ASU. I would like to fix the first protein chain and build the second protein chain with mr_rosetta. I have obtained fragment files for both proteins from robetta. What input parameters are required to tell mr_rosetta to build the second chain only? I have tried the fixed_ensemble_ID_list = ensemble1 option. Here is an excerpt of my parameter file with the input files: run a) search_models = "path/chainA.pdb" (from Phaser) search_models = "path/chainB.pdb" (homologue structure) fixed_ensembles { fixed_ensembleID_list = ensemble1 fixed_euler_list = 0.0 0.0 0.0 fixed_frac_list = 0.0 0.0 0.0 fixed_frac_list_is_fractional = True } alignment_files = "path/chainA.dat" alignment_files = "path/chainB.dat" with chainA.dat having twice the same sequence and chainB.dat with the target and template alignment. I have set fragment files with chain_list for both chains. I get this error when running phenix.mr_rosetta (running from command line with .eff file) "Sorry, the file /path/GROUP_OF_PLACE_MODEL_1/RUN_2/chainB_al_0.pdb representing group 0 has no atoms? This could indicate a problem with sequence alignment in rebuilding. It could be that there are near-duplicate sequences in your sequence file ...if this is the case then you may need to artificially rename your duplicated chains and renumber residues to make a single chain containing a duplication" This error also occurs run b) Without any alignment files run c) Without any alignment files and with just chainA.pdb as search model. An alignment of the sequences from chainA and chainB gives low identity (no duplicate regions), so I suspect the problem is located somewhere else? I have seen a post by Tom Terwilliger, explaining how to deal with two identical chains with different conformations where no alignment files are required. However, I would like to use the information from the homologue structure, that is why I specified the alignment file. By inputting chainB.pdb as search model, mr_rosetta also tries to use this model as a search model for molecular replacement, but I think this is not necessary (and gives a warning that the space group of the search model and the data file do not match). I just want to provide the structural information from structure chainB.pdb to mr_rosetta for building. ----- So in summary this is what I would like to do: chain A: keep fixed - use chainA.pdb as fixed model chain B: build - use information from homologue structure (chainB.pdb) and fragment files to build chain What is the best strategy to do this? What input parameters are required so that I do not get the error described above? ----- Thanks a lot in advance, Tobias.

3 2

phaser output and packing
by Ursula Schulze-Gahmen 26 Feb '13

26 Feb '13

Could somebody explain to me how phaser decides if there is a solution. When I look at the log file there are a few solutions in the translation function that have fewer than the allowed clashes, but they are still not recognized as potential solutions. Why is that. Is there another parameter that I should be looking at? Ursula -- Ursula Schulze-Gahmen, Ph.D. Assistant Researcher UC Berkeley, QB3 356 Stanley Hall #3220 Berkeley, CA 94720-3220

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WCPCW 21
by Williams, John Charles 22 Feb '13

22 Feb '13

All, We will be finalizing the program for the workshop on Monday, Feb 25th. The Monterey Plaza and Spa has confirmed that we can accommodate a few more people. So, if you are interested in attending and would like to be considered for an oral presentation or would like to present a poster, please register and submit an abstract no later than Monday, 12 noon (PST). The meeting will be March 17th-20th with Wayne Hendrickson as the Keystone speaker! Best, John wcpcw.org --------------------------------------------------------------------- *SECURITY/CONFIDENTIALITY WARNING: This message and any attachments are intended solely for the individual or entity to which they are addressed. This communication may contain information that is privileged, confidential, or exempt from disclosure under applicable law (e.g., personal health information, research data, financial information). Because this e-mail has been sent without encryption, individuals other than the intended recipient may be able to view the information, forward it to others or tamper with the information without the knowledge or consent of the sender. If you are not the intended recipient, or the employee or person responsible for delivering the message to the intended recipient, any dissemination, distribution or copying of the communication is strictly prohibited. If you received the communication in error, please notify the sender immediately by replying to this message and deleting the message and any accompanying files from your system. If, due to the security risks, you do not wish to receive further communications via e-mail, please reply to this message and inform the sender that you do not wish to receive further e-mail from the sender. (fpc5p) ---------------------------------------------------------------------

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Maximum Entropy Maps
by Morten Groftehauge 22 Feb '13

22 Feb '13

Hi guys, I see that phenix.maximum_entropy_map is now a command in Phenix. Some quick questions: Where is this likely to be must useful and does it take ridiculously long to run? From the Nishibori 2008 paper in Acta D it seems like this would mainly be useful for very high resolution structures that you would normally call complete - and that it would take a very long time to compute. I love trying out new methods but I need a bit more to go on with this one. Cheers, Morten -- Morten K Grøftehauge, PhD Pohl Group Durham University

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Building sugars
by Folmer Fredslund 21 Feb '13

21 Feb '13

Dear all, What's the "correct" way to build and refine sugar polymers? I am currently building several structures with different kinds of sugar polymers bound to them. Searching for similar "ligands" in the PDB, I end up with e.g. trisaccharides that are named as one molecule, even though they are indeed made up of three individual sugars with bonds between them. Thank you for any pointers. Best regards, Folmer -- Folmer Fredslund

1 0

phenix.table_one citation
by Alexander Batyuk 20 Feb '13

20 Feb '13

Dear developers, How to cite phenix.table_one? Best wishes, Alex -- Alex Batyuk The Plueckthun Lab www.bioc.uzh.ch/plueckthun

2 1

cut out density vs FFT
by Yunji Wu 19 Feb '13

19 Feb '13

Hi all-- I'm trying to make a 2Fo-Fc map with density wrapped around my asymmetric unit to make a figure showing density for my sub-8Å structure. I am not building into this map or doing MR. My model was refined in Phenix.refine (R's in the high 30's), and so I have the output model .pdb and the output map coefficients .mtz. When I input these into cut_out_density, the only "map labels" input options it gives me are 2FOFCWT, PH2FOFCWT, which seem fine. Using either box, sphere, or model for the cutout type, I get a VERY nice looking map that looks extremely solvent flattened and tight around my model, albeit still to 8Å. Suspicious of anything that looks good, I went to try the same thing in FFT (CCP4). There I generated a simple map (using the same .mtz) to cover all atoms in a pdb file- the same model I put into cut_out_density. For the input I selected 2FOFCWT and PH2FOFCWT, leaving Sigma and Weight unassigned. The map that comes out of this looks terrible, with a lot of noise and little density corresponding to the model aside from what one might expect from a very poor resolution experimental map. I've also tried using FFT in Phenix to do the same thing, and it gives like better maps than CCP4 FFT but worse than cut_out_density. It almost appears as if there is some additional solvent flattening going on, which I know happens at the beginning of refinement, but that should be intrinsic to the map coefficients file which is common to everything I've tried. My questions are (1) what should I use for what I am trying to do (just display 2Fo-Fc density around asymmetric unit for a figure) and (2) what is the difference between what these three programs (cut_out, FFT-CCP4, FFT-Phenix) are calculating? The inputs are exactly the same -- mtz (2FOFCWT and PH2FOFCWT) and pdb output from refinement. Thanks for the help, Yunji

3 2

Problems placing second component in Phaser
by Morten Groftehauge 19 Feb '13

19 Feb '13

Hi guys, So I have a solution that covers some of target and I ran it through phenix.mr_rosetta and I am actually kinda happy with the output from there. However, while I can place both (NCS=2) with the original model I only seem to be able to place one of the rosetta models, even with 'packing function' set to 'all' or 'allow'. Original model - Refinement after placing component 1: LLG 175 Refinement after placing component 2: LLG 620 Rosetta model (packing 'all') - Refinement after placing component 1: LLG 295 Refinement after placing component 2: LLG 390 Rosetta model (packing standard) - Refinement after placing component 1: LLG 295 Refinement after placing component 2: LLG 350 More notably there is little to no increase in TFZ with the placement of the second component... There are more residues in the Rosetta model and I would expect more clashes but this looks weird. Cheers, Morten -- Morten K Grøftehauge, PhD Pohl Group Durham University

3 3

where is brute rotation output
by Ursula Schulze-Gahmen 18 Feb '13

18 Feb '13

Hi, Has anybody experience with the output from the brute rotation function. I ran this and it finished without complaints, but there are no output files ***.rlist or ***.sol in the directory. I cannot figure out why and how to continue now with the translation function. From the log file it seemed that there were peaks that could have been written out. Thanks for any help Ursula -- Ursula Schulze-Gahmen, Ph.D. Assistant Researcher UC Berkeley, QB3 356 Stanley Hall #3220 Berkeley, CA 94720-3220

3 8

Phaser problem for flexible domain of protein
by Appu kumar 18 Feb '13

18 Feb '13

Dear user, I have problem in finding the phase for the flexible DNA binding domain of LTTR protein. The unit cell parameters are ,Unit Cell: 46.92 105.30 47.75 90.00 103.26 90.00, and space-Group P 1 21 1. Molecular weight of protein is 36000Da. When i am running phaser it showing that Most probable MW of protein in asu for resolution = 49714.8. Could you please guide me how to set the proper setting for finding the correct rotation function and translation function setting to get the phases for the full length protein. I m getting the phases for the ligand binding domain but not for the DNA binding domain. Could you please suggest me some tricks which would work to get the phase. Thanks a lot in advance. Appu

1 0

Trouble with Phenix
by Rachel North 18 Feb '13

18 Feb '13

Hi, I have been having some issues with Phenix. When I input the mtz file (generated from CCP4), an error message appears... "File format error: cctbx Error: invalid combination of values while extracting anomalous array from columns: F New, SIGF New, DANO New, SIGDANO New, hkl= -3,25,44." This happens when I try to use the reflection file editor, xtriage and phenix refine. I have tested datasets collected from other crystals in the same condition and still have the same problem. I have also tried to process my data using different programs (XDSME and iMOSFLM) before going to CCP4 and then Phenix for refinement. I have never had this problem before and I have run every program on CCP4 the same way that I usually do. Does anyone know how I fix this or where I am going wrong? Kind Regards

2 1

how to make covelent link of BME with Cys and refine
by Rajeshwer Sankhala 15 Feb '13

15 Feb '13

Hello Nat, Thank you so much for your suggestion. I did run phenix refinement in nightly build phenix version with " intra_chain = True " command. This time refinement worked but again in out put pdb no disulfide bond is estabilished between cys and BME. I provided .pdb, .mtz and .cif files as input. Well can you please suggest me how i can provide both BME and Cys-BME (SS bond) restrains in .cif file? if that happens then i feel refinement should work. I am attaching my .cis file below please have a look and tell me if anything wrong with this? BME.cif # electronic Ligand Builder and Optimisation Workbench (eLBOW) # - a module of PHENIX version 1.8.1-1168 (Mon Sep 28 16:48:00 2012) # - file written: Sat Jan 26 17:25:31 2013 # # Input file: Ref226_154-331-VLD2-BME.pdb # Random seed: 3628800 # Residue: BME # data_comp_list loop_ _chem_comp.id _chem_comp.three_letter_code _chem_comp.name _chem_comp.group _chem_comp.number_atoms_all _chem_comp.number_atoms_nh _chem_comp.desc_level BME BME 'Unknown ' ligand 10 4 . # data_comp_BME # loop_ _chem_comp_atom.comp_id _chem_comp_atom.atom_id _chem_comp_atom.type_symbol _chem_comp_atom.type_energy _chem_comp_atom.partial_charge _chem_comp_atom.x _chem_comp_atom.y _chem_comp_atom.z BME S S S2 . -19.3229 -20.3986 17.2400 BME H5 H H . -18.1988 -20.0550 17.0819 BME C1 C CH2 . -19.3558 -21.8852 18.1928 BME H3 H HCH2 . -18.8480 -21.7572 19.0093 BME H4 H HCH2 . -18.9637 -22.6079 17.6781 BME C C CH2 . -20.7668 -22.2261 18.5333 BME H1 H HCH2 . -21.1200 -21.5643 19.1482 BME H2 H HCH2 . -20.7965 -23.1021 18.9488 BME O O OH1 . -21.5280 -22.2370 17.3822 BME H H HOH1 . -22.3435 -22.5247 17.5704 # loop_ _chem_comp_bond.comp_id _chem_comp_bond.atom_id_1 _chem_comp_bond.atom_id_2 _chem_comp_bond.type _chem_comp_bond.value_dist _chem_comp_bond.value_dist_esd BME S H5 single 1.186 0.020 BME S C1 single 1.766 0.020 BME C1 H3 single 0.970 0.020 BME C1 H4 single 0.970 0.020 BME C1 C single 1.491 0.020 BME C H1 single 0.970 0.020 BME C H2 single 0.970 0.020 BME C O single 1.380 0.020 BME O H single 0.885 0.020 # loop_ _chem_comp_angle.comp_id _chem_comp_angle.atom_id_1 _chem_comp_angle.atom_id_2 _chem_comp_angle.atom_id_3 _chem_comp_angle.value_angle _chem_comp_angle.value_angle_esd BME C1 S H5 109.48 3.000 BME C C1 H4 109.47 3.000 BME C C1 H3 109.47 3.000 BME H4 C1 H3 109.47 3.000 BME C C1 S 109.47 3.000 BME H4 C1 S 109.47 3.000 BME H3 C1 S 109.47 3.000 BME O C H2 109.47 3.000 BME O C H1 109.47 3.000 BME H2 C H1 109.47 3.000 BME O C C1 109.47 3.000 BME H2 C C1 109.47 3.000 BME H1 C C1 109.47 3.000 BME H O C 109.48 3.000 # loop_ _chem_comp_tor.comp_id _chem_comp_tor.id _chem_comp_tor.atom_id_1 _chem_comp_tor.atom_id_2 _chem_comp_tor.atom_id_3 _chem_comp_tor.atom_id_4 _chem_comp_tor.value_angle _chem_comp_tor.value_angle_esd _chem_comp_tor.period BME Var_01 O C C1 S 52.25 30.0 3 BME Var_02 H1 C C1 S -67.75 30.0 3 BME Var_03 H2 C C1 S 172.25 30.0 3 BME Var_04 O C C1 H3 172.25 30.0 3 BME Var_05 O C C1 H4 -67.75 30.0 3 # loop_ _chem_link.id _chem_link.comp_id_1 _chem_link.mod_id_1 _chem_link.group_comp_1 _chem_link.comp_id_2 _chem_link.mod_id_2 _chem_link.group_comp_2 _chem_link.name CYS-BME CYS DEL-HG . BME DEL-HS . SS-bridge # Thanks

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