- phenixbb - phenix-online.org

March 15, 2010 deadline- User proposal submission for Collaborative Crystallography at BCSB
by Banumathi Sankaran 11 Mar '10

11 Mar '10

Dear Users, The deadline for May/June 2010 Collaborative Crystallography proposals will be *Mar 15, 2010. * ------------------------------------------------------------------------ Through the Collaborative Crystallography Program (CC) at the Advanced Light Source (ALS), scientists can send protein crystals to Berkeley Center for Structural Biology (BCSB) staff researchers for data collection and analysis. The CC Program can provide a number of benefits to researchers: * Obtain high quality data and analysis through collaborating with expert beamline researchers; * Rapid turn around on projects; and * Reduced travel costs. To apply, please submit a proposal through the ALS General User proposal review process for beamtime allocation. Proposals are reviewed and ranked by the Proposal Study Panel, and beamtime is allocated accordingly. BCSB staff schedule the CC projects on Beamlines 5.0.1 and 5.0.2 to fit into the available resources. Only non-proprietary projects will be accepted. As a condition of participation, BCSB staff researchers who participate in data collection and/or analysis must be appropriately acknowledged - typically being included as authors on publications and in PDB depositions. Please consult the website for additional information at: http://bcsb.als.lbl.gov/wiki/index.php/Collaborative_Crystallography ----------------------------------------------------------------------------------------- How To Apply: To learn more, go to: http://www.als.lbl.gov/als/quickguide/becomealsuser.html To submit a proposal, go to: http://alsusweb.lbl.gov/. Scroll down to "*Structural Biology beamlines (includes protein SAXS)*" and click on "New Proposal." Enter your proposal information, with attention to the following details: * For question 3/First choice, select "5.0.1 (Monochromatic);" for question 3/Second choice, select "5.0.2 (MAD)." * Check box (yes) in response to question (7) "Do you want collaborative crystallography (beamline 5.0.1 or 5.0.2 only) * In question 4, please describe a specific research project with a clear end point. In order to request CC time for May/June 2010 allocation period, proposals must be submitted by *March 15, 2010.* The deadline for CC proposals for the time period July/August 2010 will be May 15, 2010. Regards, Banumathi Sankaran

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does phenix.refine ignore F-obs, SIGF-obs?
by Scott Classen 11 Mar '10

11 Mar '10

Hi All, I have an mtz file that has: I-obs SIGI-obs F-obs SIGF-obs I want phenix.refine to use F-obs and SIGF-obs, but even though I provide : labels = "F-obs,SIGF-obs" I think phenix.refine is seeing the I-obs,SIGI-obs and recalculating the structure factors from those values instead of using the Fs that I provide in the mtz. Is this the case? Other than stripping the I-obs,SIGI-obs from the input mtz, is there a way to force phenix to use my structure factors? Thanks, Scott ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Scott Classen, Ph.D. SIBYLS Beamline 12.3.1 http://bl1231.als.lbl.gov Advanced Light Source Lawrence Berkeley National Laboratory 1 Cyclotron Rd MS6R2100 Berkeley, CA 94720 C) 510.206.4418 O) 510.495.2697 Beamline) 510.495.2134 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

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phenix gui freezes
by John Berrisford 11 Mar '10

11 Mar '10

Dear All A little while ago I posted a message about the GUI freezing on linux systems, either run remotely (centos) or locally (ubuntu system). I never received a solution and I'm still having this problem despite upgrading to version 1.6. On my ubuntu system the GUI seems to freeze either when the screen saver comes on (set to turn the screen off) or I switch desktop - although not everytime I do this. The freezing GUI means that I have to force quit phenix after every refinement run (I haven't tried other tasks) and that the freeR is not reported to the GUI (not a major problem). Thankfully in version 1.6 the configuration and results for each refinement run are recorded and I can retrieve this information by loading a job through job history - a big improvement over version 1.5. Anyone else having this problem and does anyone know a solution? Thanks John -- John Berrisford Medical Research Council Mitochondrial Biology Unit Wellcome Trust / MRC Building Hills Road Cambridge CB2 0XY WEB: www.mrc-mbu.cam.ac.uk TEL: +44 (01223) 252918

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Re: [phenixbb] does phenix.refine ignore F-obs, SIGF-obs?
by Scott Classen 11 Mar '10

11 Mar '10

Here are some excerpts from the phenix.refine log file: Resolution range: 44.4024 1.8604 Completeness in resolution range: 0.995991 Completeness with d_max=infinity: 0.995862 Number of F-obs in resolution range: 92411 Number of F-obs <= 0: 3322 Refinement resolution range: d_max = 44.4024 d_min = 1.8604 Fobs statistics after all cutoffs applied: Miller array info: None Observation type: xray.amplitude Type of data: double, size=89089 Type of sigmas: double, size=89089 Number of Miller indices: 89089 Anomalous flag: False Systematic absences: 0 Centric reflections: 11672 Resolution range: 44.4024 1.8604 Completeness in resolution range: 0.960187 Completeness with d_max=infinity: 0.960063 This implies to me that phenix.refine is throwing out my very weak reflections. It appears that completeness goes from 99.6% to 96.0% Then further down in the log file I see: ============================== Outliers rejection ============================= Distribution of F-obs values before outliers rejection: 0.126 - 28.714 : 65519 28.714 - 57.302 : 13850 57.302 - 85.890 : 5625 85.890 - 114.478 : 2515 114.478 - 143.066 : 991 143.066 - 171.654 : 393 171.654 - 200.242 : 128 200.242 - 228.830 : 49 228.830 - 257.418 : 12 257.418 - 286.006 : 7 basic_wilson_outliers = 0 extreme_wilson_outliers = 0 beamstop_shadow_outliers = 0 model_based_outliers = 1 total = 1 Distribution of F-obs values after outliers rejection: 0.126 - 28.714 : 65519 28.714 - 57.302 : 13850 57.302 - 85.890 : 5624 85.890 - 114.478 : 2515 114.478 - 143.066 : 991 143.066 - 171.654 : 393 171.654 - 200.242 : 128 200.242 - 228.830 : 49 228.830 - 257.418 : 12 257.418 - 286.006 : 7 which makes it look like nothing is being rejected? So what is it? in the header of the resulting pdb file: REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.860 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 44.402 REMARK 3 MIN(FOBS/SIGMA_FOBS) : 0.03 REMARK 3 COMPLETENESS FOR RANGE (%) : 96.02 REMARK 3 NUMBER OF REFLECTIONS : 89088 which implies that indeed some reflections are being rejected. My hunch is that if F-obs = SIGF-obs then the reflection is rejected. Thanks, Scott

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Re: [phenixbb] phenixbb Digest, Vol 52, Issue 12
by Kay Diederichs 11 Mar '10

11 Mar '10

Am 11.03.2010 23:51, schrieb phenixbb-request(a)phenix-online.org: > Message: 4 > Date: Wed, 10 Mar 2010 16:52:35 -0500 > From: Nathaniel Echols<NEchols(a)lbl.gov> > To: PHENIX user mailing list<phenixbb(a)phenix-online.org> > Subject: Re: [phenixbb] autosol interface bug? selinux conflict? > Message-ID:<6BE2BD93-F43C-4D37-AD91-9F31BCE2C6F1(a)lbl.gov> > Content-Type: text/plain; charset=us-ascii > > On Mar 10, 2010, at 4:37 PM, Kay Diederichs wrote: >> > I wonder >> > - why do Fortran binaries give a SElinux problem, and C++ binaries don't? > Because we bundle the Intel Fortran shared libraries (which are required by SOLVE) with the binary installers. > if you simply link with -static-intel then you don't have to distribute those libraries. HTH, Kay

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Additional residue info in coot
by Brown, Breann 11 Mar '10

11 Mar '10

Hello, I am currently refining a model of a zinc-binding protein and after refinement there is additional positive Fo-Fc density at the zinc site. This is the fourth independent structure of this particular protein solved in my lab so we are confident the metal is in fact zinc. I noticed that after refinement the "residue info" in Coot has two lines for the zinc atom. The first line reads "G/121 ZN/ZN 1.00 44.30" while the second line reads G/121 ZN/ 0.00 0.00. The occupancy for the second line cannot be changed and it only appears after refinement. I carried out the same refinement using a previously solved structure of the same protein and the extra residue info line was not present. My question is where does this extra line in the residue info come from and could the zero occupancy be causing additional Fo-Fc density at the metal site? Thanks, Breann Breann L. Brown Page Laboratory Department of Molecular Pharmacology, Physiology and Biotechnology Brown University Providence, RI 02912

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Re: [phenixbb] autosol interface bug? selinux conflict?
by Kay Diederichs 10 Mar '10

10 Mar '10

Am 11.03.2010 07:00, schrieb phenixbb-request(a)phenix-online.org: > Message: 1 > Date: Wed, 10 Mar 2010 09:28:36 -0500 > From: Nathaniel Echols<NEchols(a)lbl.gov> > To: PHENIX user mailing list<phenixbb(a)phenix-online.org> > Subject: Re: [phenixbb] autosol interface bug? selinux conflict? > Message-ID:<E0B9BF9F-C96B-4F45-9D02-CC16197AFFF0(a)lbl.gov> > Content-Type: text/plain; charset=us-ascii > > On Mar 9, 2010, at 6:03 PM, Stuart Endo-Streeter wrote: >> After this repetitive route got autosol running, I ran into some kind of >> selinux security warning that blocked autosol and caused it to crash. I >> seem to have fixed that issue by switching temporarily to permissive mode, >> but disabling security to run phenix hardly seems the appropriate solution. >> I didn't manage to catch the autosol error message, I will try to recreate >> it later tonight if needed. > > SELinux breaks the Fortran binaries we distribute. There are two things to try: > > 1. If you installed to /usr/local, try installing to your home directory instead (if this is your computer rather than a shared system). For example: > > computer:~> ./install --prefix=/home/nat/software > > I don't know if this will help or not, but it's worth a try. I wonder - why do Fortran binaries give a SElinux problem, and C++ binaries don't? - does the phenix installer script treat (i.e. label) C++ binaries in any special way which cannot be done for other binaries? If it is known that SElinux is the problem, then this already half of the solution. In general, these problems can easily be found and fixed (on RedHat-derived distros at least) by labelling libraries and binaries in a specific way which is suggested by the output of the SEtroubleshoot- daemon (to be found in /var/log/messages). If that daemon doesn't run, you may have to install it with "yum install setroubleshoot-server". HTH, Kay

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python issues
by Patrick Loll 10 Mar '10

10 Mar '10

Hi, This is really a ccp4-provoked question, but it relates to phaser & cctbx (I think), so I'll try here as well: After an install of ccp4 on a MacBook Pro running Snow Leopard and on a MacPro running Leopard, the command: source /sw*/bin/init.csh (where /sw* is either /sw64 on the machine running 10.6, or /sw on the machine running 10.5) gives the error: ******************************************* Fatal Error: Incomplete libtbx environment! ******************************************* Please re-run the libtbx/configure.py command. If I go to ...ccp4.../src/phaser/phaser-2.1.4/libtbx/ and type python configure.py Then it tells me that the md5 module is deprecated, and that the cool kids use hashlib instead. Does this mean that I need to go to an earlier version of python? Will that mess up other things? Curiously, one machines giving this problem has python 2.5.1, while the other has 2.6. However, I currently have python 2.5.1 running on another machine (albeit with a slightly earlier version of ccp4) and it doesn't give me this problem. Ideas? Thanks, Pat --------------------------------------------------------------------------------------- Patrick J. Loll, Ph. D. Professor of Biochemistry & Molecular Biology Director, Biochemistry Graduate Program Drexel University College of Medicine Room 10-102 New College Building 245 N. 15th St., Mailstop 497 Philadelphia, PA 19102-1192 USA (215) 762-7706 pat.loll(a)drexelmed.edu

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Re: [phenixbb] autosol interface bug? selinux conflict?
by Nathaniel Echols 10 Mar '10

10 Mar '10

On Mar 9, 2010, at 6:03 PM, Stuart Endo-Streeter wrote: > Problem is, column labels are selected. "i_obs sigma" is showing in the > label field for each dataset. Clicking on the field to try to re-select > makes no difference, I still get the same error message. If I load just a > single anomalous dataset in addition to the native dataset, usually autosol > will run, sometimes not. It does not matter which anomalous dataset I am > using. If I load the native and only one of the anomalous datasets it > usually works. If I then repeat the process of adding a single anomalous > dataset, running autosol, aborting the run, then adding the next dataset, > eventually I can add all the datasets and start autosol. I'm not sure what to suggest, because I can't reproduce this (with 1.6 or the latest code), nor could I find anything in the code that indicates how it happens. Could you please send me a screenshot of the GUI before you click Run? > After this repetitive route got autosol running, I ran into some kind of > selinux security warning that blocked autosol and caused it to crash. I > seem to have fixed that issue by switching temporarily to permissive mode, > but disabling security to run phenix hardly seems the appropriate solution. > I didn't manage to catch the autosol error message, I will try to recreate > it later tonight if needed. SELinux breaks the Fortran binaries we distribute. There are two things to try: 1. If you installed to /usr/local, try installing to your home directory instead (if this is your computer rather than a shared system). For example: computer:~> ./install --prefix=/home/nat/software I don't know if this will help or not, but it's worth a try. 2. Install from source. This will take several hours, so if you have multiple cores we recommend adding the flag "--nproc=8" (or whatever) when you run the install script. If you have access to the Intel Fortran compiler we recommend installing and configuring that (I think the build script will switch to using it automatically), since g77/gfortran produce binaries that are *much* slower. This problem should go away in a few months when we purge the last FORTRAN code from PHENIX. -------------------- Nathaniel Echols Lawrence Berkeley Lab 510-486-5136 NEchols(a)lbl.gov

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autosol interface bug? selinux conflict?
by Stuart Endo-Streeter 09 Mar '10

09 Mar '10

Dear board... I am trying to run autosol with a native dataset and a mad experiment to extend the phases. Unfortunately I cannot even begin the actual process (without some weird convolutions) because phenix keeps giving the following message: "You must select a column label to use in each reflection file. If no labels are shown in the drop-down menu, it may not be a valid format." Problem is, column labels are selected. "i_obs sigma" is showing in the label field for each dataset. Clicking on the field to try to re-select makes no difference, I still get the same error message. If I load just a single anomalous dataset in addition to the native dataset, usually autosol will run, sometimes not. It does not matter which anomalous dataset I am using. If I load the native and only one of the anomalous datasets it usually works. If I then repeat the process of adding a single anomalous dataset, running autosol, aborting the run, then adding the next dataset, eventually I can add all the datasets and start autosol. After this repetitive route got autosol running, I ran into some kind of selinux security warning that blocked autosol and caused it to crash. I seem to have fixed that issue by switching temporarily to permissive mode, but disabling security to run phenix hardly seems the appropriate solution. I didn't manage to catch the autosol error message, I will try to recreate it later tonight if needed. My system is CentOS 5.4, gui is KDE, phenix is v1.6, installed just yesterday. Any suggestions/clues would be appreciated. Stuart Endo-Streeter ________________________________ Stuart Endo-Streeter, Ph.D Dept. Computer Science Duke University Medical Center 3245 French Family Sciences Building 919-660-4018 stuart.endostreeter(a)duke.edu

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anisotropic refinement water hydrogens and gui
by Christian Roth 09 Mar '10

09 Mar '10

Dear all, I did an anisotropic refinement with the gui for a 1.6 Ang. structure I think this is quite O.K. as the R-values seems to be reasonable(14/18.8). But for comparison I want exclude waters and hydrogens explicitly from the anistropic refinement. One could do this in the command line mode via adp.individual.anisotropic ="not water and not element H. I started from the gui and modified the .eff directly by icnluding: adp { individual { isotropic = None anisotropic = "not water and not element H" In the output pdb all waters have an anisou flag. So I assume phenix refined them anisotropically. How do I have to modify the .eff file to exclude the waters from the anisotropic refinement. I would like to use the gui, because you get this nice summary output with all statistics. Phenix started optionally anisotropic refinement at 1.7 Ang. As far as I understand one should keep in mind that the data to parameter ration should have certain value of app. 3 or 4. But restraints as paramters are not as solid as experimental data. Is this 1.7 Ang. borderline a rule of thumb, is there a strict rule at which point I could refine anisotropic, or maybe mixed isotropic anisotropic. Best regards Christian

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phenix.refine and multi-metals centers
by Carlos Frazao 09 Mar '10

09 Mar '10

Hi I have a doubt on the refinement of e doubled metal center. I know that: 1) one may fix the metals to their actual positions by using e.g. refine.sites.individual="not element Fe" 2) one may use "phenix.metal_coordination my.pdb" to obtain a dictionary for the actual metals coordinations but how does phenix.refine handels metal-to-metal interactions? How should one set the refinement of a metallo protein with a two metals centre, in order not to bias the final result (metal to metal distances) ? Thanks a lot, Carlos -- ************************************** Dr. Carlos Frazao Structural Biology Laboratory - Macromolecular Crystallography Unit ITQB-UNL, Av Republica, Apartado 127 2781-901 Oeiras, Portugal Phone: (351)-214469666 FAX: (351)-214433644 e-mail: frazao(a)itqb.unl.pt www.itqb.unl.pt

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molprobity todo list and coot integration in linux
by hari jayaram 09 Mar '10

09 Mar '10

Hi, Is a difference in the way the GUI molprobity TODO list is displayed between linux and Mac OSX . I am running 1.6.289 on both ...and have coot with scheme and python scripting installed, On mac OSX , phenix starts up coot with the molecule and maps loaded after a phenix.validate and a TO-DO list in a separate GUI window In Linux these separate To-do list windows dont seem to open . ..I have looked everywhere and the dont seem burried behind something else, Is there some setting I need to toggle to have these popup , sortable todo lists for molprobity assisted rebuilding on Linux systems? Thanks for your help Hari

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Autosol + MR
by Manoj saxena 07 Mar '10

07 Mar '10

Hi, I have partial solution from MR on a crystal with dimensions 52.729 60.364 142.908 90.00 90.00 90.00 P 21 21 21. I have SAD data from another crystal with dimension 53.860 61.470 142.492 90.00 90.00 90.00 P 21 21 21. My question is can I treat both as isomorphous and combine data from both of them and use my MR as model with Autosol. What is the maximum difference in unit cell dimensions allowed to consider two unit cell isomorphous? If you can point to some references/examples that will be highly appreciated. Thanks a lot. Marusa

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phenix.reduce segmentation fault with -trim option on Snow leopard Mac OSX
by hari jayaram 04 Mar '10

04 Mar '10

Hi I am running the phenix.reduce with the -trim option and getting a segmentation fault with phenix version 1.6.289 I will remove the molprobity introduced hydrogens using pdbtools ..but thought I will write in with this Thanks Hari $ phenix.reduce s3-3_2.1_refmac0-coot-15_refmac0FH.pdb -trim > s3-3_2.1_refmac0-coot-15_refmac0FnoH.pdb reduce: version 3.15 11/6/2009, Copyright 1997-2009, J. Michael Word Processing file: "s3-3_2.1_refmac0-coot-15_refmac0FH.pdb" Segmentation fault

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coot integration require coot with python?
by hari jayaram 03 Mar '10

03 Mar '10

Hi Is it likely that the phenix-coot integration with the new GUI requires a python infused coot ?. Although I have set the coot path correctly nothing happens after coot launches . I am using the ccp4 6.1.3 coot 0.6 (revision 2540) [with guile 1.8.3 embedded] and phenix 1.6.289 on OSX snow leopard 10.6.2. I know the ccp4 coot does not come with python scripting support ...the pymol integration seems fine , but coot just sits there with an empty window after a phenix.validate run. Any ideas on how to have them talking to each other? Thanks Hari

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autosol and NCS
by hli＠uci.edu 03 Mar '10

03 Mar '10

I have solved a structure by MR with Phaser. There are 4 copies of molecules per ASU. Parts of the search model are not agreeable with the new structure leading to messy density in some regions. Now a set of SAD data has been collected. I want to run Autosol (MRSAD) to combine the SAD and model phases, doing density modification including the NCS. Can I run simple_ncs_from_pdb to get the NCS operators from the MR solution pdb file and use them for Autosol? Thanks for help. Huiying UC Irvine

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Phaser phasing power statistics
by Daqi Tu 03 Mar '10

03 Mar '10

Hi, I was trying to find phasing power statistics from phenix.autosol log file but couldn't. My structure was phased by Phaser SAD module in phenix.autosol, all I could find was figure of merit. I then ran Phaser SAD within CCP4 and still could not find phasing power statistics from its log. Does anyone know if Phaser SAD generates phasing power statistics in its log file at all? Thanks. Daqi Tu Postdoc Eck lab Harvard medical school

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SAD + Xray = ???
by J. Fleming 02 Mar '10

02 Mar '10

Hi, I have a set of Se-Met SAD data, high resolution x-ray data, and a protein sequence. Is there a way to combine these data sets in Phenix and carry out an autobuild? I saw a section in the manual about MRSAD but that requires a starting model. I've used the SAD data alone to generate a model but I'd like to use both data sets together. Thanks for your thoughts, -Jon

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ICCBM13 - Call for abstracts now open!
by Valerie Pye 02 Mar '10

02 Mar '10

Apologies for this being off topic - but may be of general interest: * * * * *13th International Conference on the Crystallisation of Biological Macromolecules* Trinity College Dublin, Ireland 12th – 16th September 2010 *Crystallization Workshop* – 9th – 12th September 2010 Call for Abstracts Now Open <http://www.iccbm13.ie/abstracts.html> The *13th International Conference on the Crystallisation of Biological Macromolecules* (*ICCBM13 <http://www.iccbm13.ie/>*) is the meeting of choice to learn about the latest research and developments in biomacromolecular crystallogenesis for structure-function-rational design applications. At ICCBM13 leading academic and pharmaceutical scientists will report on challenging systems to include macromolecular complexes and membrane proteins with an emphasis on novel practical as well as mechanistic aspects of crystallogenesis. To cater to the needs of the individual wanting to learn how to grow crystals or to acquire new crystallisation skills, the Conference is preceded by a *Workshop <http://www.iccbm13.ie/workshop.html>* devoted to hands-on instruction on select aspects of macromolecule production and crystal growth. The Conference will be held at *Trinity College Dublin <http://www.tcd.ie/>, Ireland* from the 12th to the 16th September 2010. Convenient and affordable housing is available on and around Trinity College. The campus is situated in the heart of Ireland’s capital city and is surrounded by a diversity of restaurants, pubs, theatres and other points of interests. *Scholarships <http://www.iccbm13.ie/scholarships.html>* are available to Graduate Students and Postdoctoral Fellows. Additional information is available at *www.iccbm13.ie* *ICCBM13 Congress Secretariat* T: + 353 (0) 1 400 3626 F: + 353 (0) 1 400 3692 E: info(a)iccbm13.ie W: www.iccbm13.ie *C/O Keynote PCO* Suite 26, Anglesea House, Blackrock Co. Dublin, Ireland www.keynotepco.ie

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twin refinement
by subhash bihani 02 Mar '10

02 Mar '10

Hi i am refining a protein strucutre with space group P61. Intesity statistics doesnot suggest any twining but refmac and phenix xtriage suggest a twin operator with fraction 0.48. when refined with this twin operator R factors come dowm by 3-5% and also map looks much better. but upon closer inspceion it seems that maps are heavily biased toward the model in both refmac and phenix ( I have mutated one lysine to his and it changed the map to cover his). my question is why twin refinement maps are biased and what are the ways to overcome this. my second question is how to decide whether to use twin refinement or not. is it depends on R factor statistics or we have to check some other parameters. thanks in advance subhash bihani scientific officer Bhabha Atomic Research Centre Trombay-400085 Mumbai(M.S.) India Ph. +912594688 (o) +919322191020 -- Subhash Bihani Scientific Officer Solid State Physics Division Bhabha Atomic Research Centre Trombay, Mumbai-85 Ph. 25594688 (o) alternate email: subhashbihani(a)yahoo.co.in

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NCS restraints
by Mirek Cygler 28 Feb '10

28 Feb '10

Hi, I have NCS symmetry in my crystal and wouldlike to use it during the refinement. What is the strength of the restraints for the backbone, for the sidechains? How can I control these restraints? Thanks, Mirek

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Clarification on ADPs in PDB files for TLS refinement ?
by Phil Jeffrey 27 Feb '10

27 Feb '10

Hi, Having a discussion with a colleague yesterday on differences in Phenix and Refmac representations of TLS B-factors in the PDB file I came across two different definitions for what the PDB ANISOU U's correspond to as written by Phenix: 1. From necat.chem.cornell.edu/workshops/.../WK04_PavelAfonine.pdf on p.20 U(TOTAL)= U(ATOM) + U(TLS) + U(CRYST) and only U(atom) + U(tls) is stored in the PDB ANISOU card. 2. From http://proteincrystallography.org/ccp4bb/message8948.html "Utotal = Utls + Ulocal + Ucryst. So, the ANISOU records always contain Utotal and ATOM records contain isotropic equivalent of Utotal" Which is not the same thing. I suspect it's #1. Am I right ? I would think that U(cryst) is potentially a non-zero contribution to atomic ANISOU values but is actually applied before the TLS/Anisotropic B-factors are refined. As a tie-breaker between #1 and #2 the program documentation suggest that it is just U(tls)+U(atom) but the paragraph that nominally explains it is not short of of inconsistencies, referring to the ANISOU record as having the "the total B-factor (B_tls + B_individual)" whereas they're not only not B's (they are U's) but it's also not the *total* U or B by its own definitions. IMHO it would warrant a rewrite for clarity since B and U are used interchangeably and "total" has variable usage. Phil Jeffrey Princeton

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Re: [phenixbb] Map after refinement
by Xuewu Zhang 26 Feb '10

26 Feb '10

To Paul, No, I used the mtz format map file in coot. TO Pavel, I will send you the files tomorrow. The command I used is the simplest one: phenix.refine input.pdb input.mtz. By the way, the same thing happened to one of my colleagues here a while ago. She solved the problem by updating to the then new 1.5 version. Thanks you guys for the quick replies. Xuewu Zhang UT Southwestern Medical Center On Fri, Feb 26, 2010 at 4:29 PM, <phenixbb-request(a)phenix-online.org> wrote: > Send phenixbb mailing list submissions to > phenixbb(a)phenix-online.org > > To subscribe or unsubscribe via the World Wide Web, visit > http://phenix-online.org/mailman/listinfo/phenixbb > or, via email, send a message with subject or body 'help' to > phenixbb-request(a)phenix-online.org > > You can reach the person managing the list at > phenixbb-owner(a)phenix-online.org > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of phenixbb digest..." > > > Today's Topics: > > 1. Int'l Workshop on Macromolecular Crystallography - South > America (Alejandro Buschiazzo) > 2. Re: Gap between R and Rfree (Felix Frolow) > 3. Re: Map after refinement (Xuewu Zhang) > 4. Re: Map after refinement (Paul Adams) > 5. Re: Map after refinement (Pavel Afonine) > 6. weaker density in maps (David Garboczi) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 26 Feb 2010 18:17:06 -0200 > From: Alejandro Buschiazzo <alebus(a)pasteur.edu.uy> > To: PHENIX user mailing list <phenixbb(a)phenix-online.org> > Subject: [phenixbb] Int'l Workshop on Macromolecular Crystallography - > South America > Message-ID: <4B882C42.9080701(a)pasteur.edu.uy> > Content-Type: text/plain; charset=windows-1252; format=flowed > > Dear colleagues, > > just a reminder, we're close to the deadline for applications. Details > below. > > Thank you! > > > ----------------------------------------------------------------------------------------------------------------------- > > *International Workshop ?Macromolecular Crystallography : Introduction > and Applications"* > > *April 26 - May 7, 2010* > > *Institut Pasteur de Montevideo* > > *Uruguay* > > The principal aim of this course, is to give intense hands-on training > in all main aspects of macromolecular 3D structure determination by X > ray diffraction. > > The course will include lectures delivered by a number of > international experts in the discipline, linked to hands-on > experimental work and demonstrations. > > Special emphasis will be given to : > > * Experiment design : manual and robotic crystallogenesis of > macromolecules, X ray diffraction of single crystals, data collection > * Diffraction data processing > * Model refinement and structural analysis > > > *ACTIVITY INTENDED FOR* > > * Graduate Students (MSc and PhD) > * Postdocs > * Young Research Scientists > > > *APPLICATION REQUIREMENTS* > > Candidates should send the following information : > > 1. -Short CV (including age, current position, scientific interests, > publications) > 2. -Letter of motivation (the applicant should stress how > Macromolecular Crystallography is included in her/his ongoing > scientific project) > 3. -Reference letter from the apllicant's supervisor > > > *SCHOLARSHIPS* > > Inscription fees are fully covered for all accepted students. > Applicants from abroad with no financial support, are welcome to apply > for a lodging and per diem expenses scholarship. This should be > inlcuded as a separate document at the time of application. A limited > number of traveling aids will be available: arguments backing this > request should be duly explained, to be evaluated on a case by case > basis. > > *APPLICATION DEADLINE* > > All applications should be received until March 7 2010. They should be > sent as a single pdf document to : crystallography(a)pasteur.edu.uy > > *ORGANIZERS* > > Dr. Alejandro Buschiazzo > Unidad de Cristalograf?a de Prote?nas > Institut Pasteur de Montevideo > > Dr. Pedro Alzari > Departamento de Biolog?a Estructural y Qu?mica > Institut Pasteur de Paris > > Dr. William Shepard > Proxima 2 > Synchrotron Soleil > > For further information, please E-mail to crystallography(a)pasteur.edu.uy > > The official language of the course will be english. > > Detailed information is available at: > http://www.pasteur.edu.uy/mxcourse Please check for regular updates of > the program. > > *FUNDING* > > RIIP, AMSUD/PASTEUR, IP MONTEVIDEO (URUGUAY INNOVA), CeBEM > > This International Workshop is part of the activities of the Center of > Structural Biology of Mercosur (Centro de Biolog?a Estructural del > Mercosur - CeBEM) > > > > > -- > Alejandro Buschiazzo, PhD > Research Scientist > Laboratory of Protein Crystallography > Pasteur Institute of Montevideo > Mataojo 2020 > Montevideo 11400 > URUGUAY > > Phone: +5982 5220910 int. 120 > Fax: +5982 5224185 > > > > ------------------------------ > > Message: 2 > Date: Fri, 26 Feb 2010 22:18:33 +0200 > From: Felix Frolow <mbfrolow(a)post.tau.ac.il> > To: PHENIX user mailing list <phenixbb(a)phenix-online.org> > Subject: Re: [phenixbb] Gap between R and Rfree > Message-ID: <F52C4FE8-AE34-4C23-B74E-A923995E5831(a)post.tau.ac.il> > Content-Type: text/plain; charset=us-ascii > > Yea Phil, You have got them > I was accused several years ago for over-fitting Rfree !!!!!!! > The bad thing is that to publish you have to avoid arguing with these > post-crystallography era referees. > Today they ask meaningless question " I am novice to crystallography, teach > me how to minimize Rfree" > Tomorrow they referee you papers and make meaningless remarks about gap > between R and Rfree > " Your R factor is 17% and Rfree is 24%, it rase question" > geeeeek!!!!!! what question????? > Dr Felix Frolow > Professor of Structural Biology and Biotechnology > Department of Molecular Microbiology > and Biotechnology > Tel Aviv University 69978, Israel > > Acta Crystallographica D, co-editor > > e-mail: mbfrolow(a)post.tau.ac.il > Tel: ++972 3640 8723 > Fax: ++972 3640 9407 > Cellular: ++972 547 459 608 > > On Feb 26, 2010, at 11:30 , Phil Evans wrote: > > > > > I do worry a bit about the fetishisation of R-factors, to the point where > I get the impression that some people think that the object of refinement is > minimisation of Rfree. The aim is to produce the "best" model consistent > with the data, and the most important tool is looking at the maps on the > graphics, in conjunction with validation tools analysing consistency with > what we know about stereochemistry (Ramachandran, rotamers, bumps etc). Also > is your model consistent with your initial experimental phased map, if > available? The Rfree is a useful guide, but no more than that. > > > > My (admittedly superficial) experience of changing weights, freeR > selection, resolution limits, NCS, etc in refinement is that often you can > spend some time doing this, maybe improve the R-factors a bit, but end up > with a model which is indistinguishable from the previous model when > superimposed on it. > > > > I'm fed up with referees telling me my structure must be wrong because > the Rfree is "too high" (and I haven't followed their particular pet > protocol), when I know that it is essentially correct, and as good as I can > make it. > > > > Phil > > > > > > On 26 Feb 2010, at 01:49, Pavel Afonine wrote: > > > >> Hi Simon, > >> > >> the next available PHENIX nightly build will have this command: > >> > >> phenix.r_factor_statistics > >> > >> This will output a plain text histograms for Rwork, Rfree and > Rfree-Rwork. For example: > >> > >> *** > >> phenix.r_factor_statistics 2.5 left_offset=0.2 right_offset=0.2 n_bins=5 > >> Command used: > >> > >> phenix.r_factor_statistics 2.500 left_offset=0.200 right_offset=0.200 > n_bins=5 > >> > >> > >> Histogram of Rwork for models in PDB at resolution 2.30-2.70 A: > >> 0.115 - 0.168 : 149 > >> 0.168 - 0.220 : 2998 > >> 0.220 - 0.273 : 1879 > >> 0.273 - 0.325 : 67 > >> 0.325 - 0.378 : 2 > >> Histogram of Rfree for models in PDB at resolution 2.30-2.70 A: > >> 0.146 - 0.210 : 116 > >> 0.210 - 0.274 : 3288 > >> 0.274 - 0.337 : 1664 > >> 0.337 - 0.401 : 26 > >> 0.401 - 0.465 : 1 > >> Histogram of Rfree-Rwork for all model in PDB at resolution 2.30-2.70 A: > >> 0.001 - 0.021 : 233 > >> 0.021 - 0.041 : 1412 > >> 0.041 - 0.060 : 2180 > >> 0.060 - 0.080 : 1045 > >> 0.080 - 0.100 : 225 > >> Number of structures considered: 5095 > >> *** > >> > >> and running it without arguments will take all structures. > >> > >> Pavel. > >> > >> > >> > >> On 2/25/10 2:01 PM, Simon Kolstoe wrote: > >>> oh yes please Pavel (command line polygon/statistics overview) - could > the output be a pdf or ps rather than just starting up the GUI? > >>> > >>> Simon > >>> > >>> > >>> > >>> On 25 Feb 2010, at 19:39, Pavel Afonine wrote: > >>> > >>>> Hi Francis, > >>>> > >>>> in PHENIX GUI: "Validation" -> "PDB Statistics Overview" > >>>> > >>>> or: > >>>> > >>>> in PHENIX GUI: "Validation" -> "POLYGON" > >>>> > >>>> The underlying idea is published here: > >>>> > >>>> Crystallographic model quality at a glance. L. Urzhumtseva, P. V. > Afonine, P. D. Adams, A. Urzhumtsev Acta Cryst. D65, 297-300 (2009) > >>>> > >>>> I can add a command line version if anyone is interested. > >>>> > >>>> Pavel. > >>>> > >>>> > >>>> > >>>> On 2/25/10 9:45 AM, Francis E Reyes wrote: > >>>>> Pavel > >>>>> > >>>>> What phenix utility produces such information? > >>>>> Thx > >>>>> > >>>>> FR > >>>>> > >>>>> On Feb 25, 2010, at 10:43 AM, Pavel Afonine <PAfonine(a)lbl.gov> > wrote: > >>>>> > >>>>>> Hi Young-Jin, > >>>>>> > >>>>>> the distribution of R-factors for all structures in PDB refined at > resolutions between 2.4 and 2.6 A is following: > >>>>>> > >>>>>> Histogram of Rwork for all model in PDB at resolution 2.40-2.60: > >>>>>> 0.115 - 0.141 : 5 > >>>>>> 0.141 - 0.168 : 69 > >>>>>> 0.168 - 0.194 : 414 > >>>>>> 0.194 - 0.220 : 955 <<<<< your structure > >>>>>> 0.220 - 0.246 : 695 > >>>>>> 0.246 - 0.273 : 153 > >>>>>> 0.273 - 0.299 : 25 > >>>>>> 0.299 - 0.325 : 5 > >>>>>> 0.325 - 0.352 : 0 > >>>>>> 0.352 - 0.378 : 1 > >>>>>> Histogram of Rfree for all model in PDB at resolution 2.40-2.60: > >>>>>> 0.146 - 0.178 : 3 > >>>>>> 0.178 - 0.210 : 41 > >>>>>> 0.210 - 0.242 : 404 > >>>>>> 0.242 - 0.274 : 1104 > >>>>>> 0.274 - 0.305 : 653 <<<<< your structure > >>>>>> 0.305 - 0.337 : 108 > >>>>>> 0.337 - 0.369 : 8 > >>>>>> 0.369 - 0.401 : 0 > >>>>>> 0.401 - 0.433 : 0 > >>>>>> 0.433 - 0.465 : 1 > >>>>>> Histogram of Rfree-Rwork for all model in PDB at resolution > 2.40-2.60: > >>>>>> 0.002 - 0.012 : 28 > >>>>>> 0.012 - 0.022 : 83 > >>>>>> 0.022 - 0.031 : 232 > >>>>>> 0.031 - 0.041 : 430 > >>>>>> 0.041 - 0.051 : 458 > >>>>>> 0.051 - 0.061 : 493 > >>>>>> 0.061 - 0.071 : 298 > >>>>>> 0.071 - 0.080 : 186 > >>>>>> 0.080 - 0.090 : 85 <<<<< your structure > >>>>>> 0.090 - 0.100 : 29 > >>>>>> > >>>>>> which I interpret as your Rwork is most likely good as well as > Rfree, but the gap Rfree-Rwork is too large meaning possible overfitting. > >>>>>> > >>>>>> A few tips: > >>>>>> - if you have NCS - use it in refinement; > >>>>>> - optimize refinement target weights: a) automatically: > "optimize_wxc=true optimize_wxu=true", or b) manually using wxc_scale and > wxu_scale parameters; > >>>>>> - make sure you use the latest PHENIX version. > >>>>>> > >>>>>> Let me now if you have any questions. > >>>>>> > >>>>>> Pavel. > >>>>>> > >>>>>> > >>>>>> On 2/25/10 9:17 AM, Young-Jin Cho wrote: > >>>>>>> Hi everyone, > >>>>>>> I recently got a diffraction data and am trying to refine it. The > question is whatever I did the gap between R and Rfree stay far away: > .2140/.2982 around 2.5 A resolution. (water added and > isotropic(individual_adp) and so on). > >>>>>>> Although I am redoing with many different approaches, if anyone can > give me any comments or suggestions, it would be helpful. > >>>>>>> > >>>>>>> Thanks in advance, > >>>>>>> > >>>>>>> Young-Jin > >>>>>>> > >>>>>>> > >>>>>>> _______________________________________________ > >>>>>>> phenixbb mailing list > >>>>>>> phenixbb(a)phenix-online.org > >>>>>>> http://phenix-online.org/mailman/listinfo/phenixbb > >>>>>>> > >>>>>> _______________________________________________ > >>>>>> phenixbb mailing list > >>>>>> phenixbb(a)phenix-online.org > >>>>>> http://phenix-online.org/mailman/listinfo/phenixbb > >>>>> _______________________________________________ > >>>>> phenixbb mailing list > >>>>> phenixbb(a)phenix-online.org > >>>>> http://phenix-online.org/mailman/listinfo/phenixbb > >>>> _______________________________________________ > >>>> phenixbb mailing list > >>>> phenixbb(a)phenix-online.org > >>>> http://phenix-online.org/mailman/listinfo/phenixbb > >>> > >>> _______________________________________________ > >>> phenixbb mailing list > >>> phenixbb(a)phenix-online.org > >>> http://phenix-online.org/mailman/listinfo/phenixbb > >> _______________________________________________ > >> phenixbb mailing list > >> phenixbb(a)phenix-online.org > >> http://phenix-online.org/mailman/listinfo/phenixbb > > > > _______________________________________________ > > phenixbb mailing list > > phenixbb(a)phenix-online.org > > http://phenix-online.org/mailman/listinfo/phenixbb > > > > ------------------------------ > > Message: 3 > Date: Fri, 26 Feb 2010 14:31:26 -0600 > From: Xuewu Zhang <xuewu.zhang(a)gmail.com> > To: phenixbb(a)phenix-online.org > Subject: Re: [phenixbb] Map after refinement > Message-ID: > <926abad51002261231g71b81d30o48b7ab63f41ba820(a)mail.gmail.com> > Content-Type: text/plain; charset="iso-8859-1" > > Hi Phenix developers/gurus, > I got a SAD structure, the experimental map (overall_best_denmod..mtz) from > Autosol looks great. I then built the model and used phenix.refine to > refine > the model using the "exptl_fobs....mtz", which worked well as shown by the > lowered Rfree. However, the map from the refinement makes no sense at all, > totally featureless. Using the same refined model and mtz, > phenix.model_vs_data did make the right map. I have tried both phenix 1.5 > and 1.6. Any clue? > Thanks, > Xuewu Zhang > UT Southwestern Medical Center >

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Re: [phenixbb] Map after refinement
by Xuewu Zhang 26 Feb '10

26 Feb '10

Hi Phenix developers/gurus, I got a SAD structure, the experimental map (overall_best_denmod..mtz) from Autosol looks great. I then built the model and used phenix.refine to refine the model using the "exptl_fobs....mtz", which worked well as shown by the lowered Rfree. However, the map from the refinement makes no sense at all, totally featureless. Using the same refined model and mtz, phenix.model_vs_data did make the right map. I have tried both phenix 1.5 and 1.6. Any clue? Thanks, Xuewu Zhang UT Southwestern Medical Center

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