June 2009 - phenixbb - phenix-online.org

Re: [phenixbb] phenix.automr ensemble/component question
by Thomas C. Terwilliger 23 Jul '10

23 Jul '10

Hi Scott, Your first command looks good to me. It is saying that the composition of the crystal is 2 copies of each chain: component_1.component_copies=2 component_2.component_copies=2 and you want to search for 2 copies of each of your search models: ensemble_1.copies_to_find=2 ensemble_2.copies_to_find=2 All the best, Tom T > Hi Phenixers, > > I'm a bit confused about the proper syntax for my phenix.automr job. > This is probably a simple concept, but I'm running on 3 hours of sleep > as my kid was up all night, and my synapses aren't working so good. > > I expect 2 copies of my protein per AU > > My protein is composed of two domains > > I have a single PDB search model for each domain > > is either of these commands correct? > > phenix.automr data=mydata.sca \ > ensemble_1.coords=domain1.pdb \ > ensemble_1.identity=95 \ > ensemble_1.copies_to_find=2 \ > component_1.mass=40579 \ > component_1.component_copies=2 \ > ensemble_2.coords=domain2.pdb \ > ensemble_2.identity=95 \ > ensemble_2.copies_to_find=2 \ > component_2.mass=13410 \ > component_2.component_copies=2 > > > or > > > phenix.automr data=mydata.sca \ > ensemble_1.coords=domain1.pdb \ > ensemble_1.identity=95 \ > ensemble_1.copies_to_find=1 \ > component_1.mass=40579 \ > component_1.component_copies=2 \ > ensemble_2.coords=domain2.pdb \ > ensemble_2.identity=95 \ > ensemble_2.copies_to_find=1 \ > component_2.mass=13410 \ > component_2.component_copies=2 > > > essentially I'm not really using an ensemble of search models, I only > have a single PDB for each domain. So should ensemble_1.copies_to_find > and ensemble_2.copies_to_find be set to 1 or 2? > > > 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 > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ > > > > > _______________________________________________ > phenixbb mailing list > phenixbb(a)phenix-online.org > http://www.phenix-online.org/mailman/listinfo/phenixbb >

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map output by phenix.refine
by Pavel Afonine 30 Jun '09

30 Jun '09

Hi Everyone, I think it's time to review this subject since it is one of the most frequently asked questions. Ironically, I think this is because we tried to make it as clear as possible -:) So, what kind of maps phenix.refine outputs by default? phenix.refine outputs two types of maximum-likelihood weighted maps (or, in other words, sigmaa-weighted maps): 2mFo-DFc and mFo-DFc. Now, as many of you noticed, the MTZ file with map coefficients "_map_coeffs.mtz" contains in fact four maps: 2mFo-DFc and mFo-DFc, and "filled" 2mFo-DFc and mFo-DFc (in fact fo-fc maps should be nearly identical; actually I have to fix it and not write identical fo-fc maps). The first two maps are computed using original Fobs (Fo), and the last two maps are computed using "filled Fobs", that is the original Fobs where missing reflections are "filled" with DFc. It is well known (I can spell a long list or references) that the data incompleteness affects the map quality, and sometimes, certain types of data incompleteness can *severely* distort maps. A possible solution (in order to reduce this negative effect) is to "model" missing Fobs somehow. One possibility is just to put in DFc in place where Fobs is missing, or as suggested by the classics, one can use <Fobs> taken in a resolution bin around a missing reflection. I even tried to use the random numbers and it was also better than doing nothing. Obviously, there is a nearly invisible line between the benefits of "filling in" missing Fobs and introducing bias. Where this line goes - is the subject of a research that to my knowledge is not done yet. Anyway, this is why phenix.refine writes out "regular" and "filled" maps: one is to give you unbiased but eventually lower quality map, and the other one is to give you a better-looking map with a risk of being biased. This way users have more options in exploring their maps (and less reasons for saying that Refmac produces better-appearing maps than phenix.refine -- see below). I have to mention that to my knowledge REFMAC always writes "filled" maps (those with missing Fobs substituted by DFc): - it is mentioned in Maria Turkenburg's thesis: http://www.ysbl.york.ac.uk/~mgwt/thesis-tth/chapter2.html#tth_sEc2.6.5 - and in Refmac docs: http://www.ccp4.ac.uk/html/refmac5/keywords/xray-general.html "Missing Data: For those reflections where the FP are missing, mFo is set equal to dFc. (...)". Correct me if I'm missing something. Please let me know if I wasn't clear in my attempt to explain this. I will update the phenix.refine documentation accordingly. Pavel.

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the different between 2mFo-DFc and mFo-DFc map
by 丁玮 30 Jun '09

30 Jun '09

hi,everybody, By phenix.refine outputde two likelihood-weighted maps: 2mFo-DFc and mFo-DFc.When they were displayed in coot, There is a vast difference between them, why? Thanks dream830106 　　　　　　　　dingding830106(a)sina.com 　　　　　　　　　　2009-06-30

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Autobuild requires more then 24 hrs to run on 32 cpu?
by Tatyana Sysoeva 30 Jun '09

30 Jun '09

Hello! I am trying to use Autobuild wizard and it seems that it takes longer time that I expected. That's why I have next questions: 1. What is "usual" time range estimate for Autobuild run with default parameters if my model has 7 chains with 284 aa in each? (if I'm running Autobuild on a single machine it does not finish in 3 days or on cluster with usage of 32 processors - in 24 hours) 2. Do I use phenix comman correctly for using several machines at once? I used /phenix-1.4-89, command line *phenix.autobuild data=e239a.hkl seq_file=seq.dat model=model.pdb nproc=32* In 24 hrs, which is my limit on cluster time, I get next things - AutoBuild_run_1_1_/AutoBuild_run_1_1.log ends with *Setting up to build 5 models to be combined into final model #1 NOTE: input model will be rebuilt in place: nothing will be added or deleted. Building 5 models ... Running up to 32 jobs in parallel... Splitting work into 5 jobs and running 32 at a time with csh in /home2/tas354/phenix_runs/20090610/1/AutoBuild_run_1_/TEMP0 Starting job 1... Starting job 2... Starting job 3... Starting job 4... Starting job 5...* AutoBuild_run_1_1_/TEMP0/AutoBuild_run_1_N.log ends with *Copying refined_pdb_in.pdb to composite_model.pdb in AutoBuild_run_1_/TEMP0 Beginning merge with current best model: refined_pdb_in.pdb Merging models: composite_model.pdb refine_1.pdb Refining model: Build_composite_3.pdb Model: AutoBuild_run_1_/TEMP0/Build_composite_refined_3.pdb R/Rfree=0.23/0.25 Rebuilding side chains from Build_composite_refined_3.pdb* or with error * TRIED resolve_extra_huge ...but not OK ******************************************************************************** Failed to carry out AutoBuild_build_cycle: ******************************************************************************** * Should I change parameters of the Autobuild which will dramatically reduce time for computations or should I ask for extension of my time for letting the programs to run as it is? I would give more details if it is needed I just could not think of some other right now. And I would appreciate any advices and suggestions! Thank you Tanya

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phenix.refine simulated annealing modes
by Alejandro Buschiazzo 29 Jun '09

29 Jun '09

Dear Phenix users, I can't find information on the precise meaning of the different modes to perform SA during refinement; some choices are self-explanatory (such as "every_macro_cycle" or "second_and_before_last"), but what about "once" or "first_half"? (in my particular case, I want to perform only one simulated annealing routine on the second macro_cycle: can I use "once"? but, where should I define at which cycle to perform it?) Thank you for your help, Best, -- 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

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questoin about AutoBuild
by crystallogrphy 27 Jun '09

27 Jun '09

Deal All, After I tried 9 runs of AutoSol from my MAD data by changing parameters (resolution, heavy atom numbers), I got 9 results, named as: AutoSol_run_1_ to AutoSol_run_9_. I found #9 and #7 have the lowest Rfree, so I want to pick the model from these two runs. However, when I use "phenix.autobuild after_autosol", the resulting AutoBuild files tell me that the program picked the result from #5 of AutoSol, not the lastest #9. Does anyone know how can I specify the exact number of AutoSol to run autobuild? I also tried this command "phenix.autobuild data=resolve_3.mtz model=overall_best.pdb seq_file=zntb-sd.seq"under the directory of AutoSol_run_9_, but failed with the following word: ******************************************************************************** Failed to carry out AutoBuild_build_cycle: Initial phenix refinement of AutoBuild_run_2_/TEMP0/NO_LIG_NO_WAT_overall_best_edited.pdb with exptl_fobs_phases_freeR_flags.mtz failed...quitting... See AutoBuild_run_2_/LAST.LOG for more information Please resolve the R-free flags mismatch. ******************************************************************************** Does anyone know where is wrong? Qun Wan Department of Pharmacology, Case Western Reserve University 10900 Euclid Avenue, Cleveland, OH 44106 email: qunwan1(a)gmail.com lab phone: 216-368-3337

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"phenix.fobs_minus_fobs_map" new tool in PHENIX
by Pavel Afonine 27 Jun '09

27 Jun '09

Hi Everyone, for all of you who asked about computing Fo-Fo map in PHENIX: I added a new command line tool called "phenix.fobs_minus_fobs_map". For usage details just type phenix.fobs_minus_fobs_map and hit Enter, and it will print a couple of examples. Any feedback will be very appreciated. This is available in development version of PHENIX starting from 1.4-51: goto: http://www.phenix-online.org/download/ then click "Latest installers" under "Nightly builds (version 1.4-4 and above)" section. Pavel.

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Refining partially occupied DNA on top of itself
by bschmidt＠berkeley.edu 26 Jun '09

26 Jun '09

Hello, I am having difficulty using Phenix to build and refine a DNA duplex. The issue is that my asymmetric unit consists of one protein monomer bound to DNA, but the protein is a dimer and the DNA is not palindromic, so each monomer is bound to a different sequence of DNA. As such, the density for the two different DNA half-sites is averaged out in the asymmetric unit. I have tried to place two duplexes directly on top of one another, each with 0.5 occupancy, and then refine. But I have noticed two problems. The first is that when xyz refinement is off, and I look at the output files, the density for DNA is awfully green, as if there were only *one* helix with 0.5 occupancy there. The other problem I noticed is that when I turn xyz refinement on, and look at the output files, one of the two half-sites gets moved several angtroms, so that it is in a region that generally has no density. I expect that, if done properly, the backbone of both half-site DNAs ought not to move. Any advice would be greatly appreciated. Thanks, Bryan Schmidt

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multi-xtal averaging vs twinning
by Frank von Delft 26 Jun '09

26 Jun '09

Hi, I'm trying to to figure out how to transfer phases from a twinned SeMet dataset to the native one for refinement. 1) SeMet dataset (2.9A) is solved (despite twinning), structure built, refines 29/35 (ish) 2) SeMet crystals are hemihedrally twinned (confirmed by refining with twin law), native probably too. 3) Native dataset (2.7A) has 5A difference in the long cell edge (478 vs 483A). So I could (and will try to) simply copy the SeMet phases into the native dataset, even though CC Fnat and Fsme is rather low (80-20%). But would phenix.multi_crystal_average know what to do with this? It has no explicit options, so I expected not. Or how *would* one approach this? Cheers Frank

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Fix for autobuild/ligandfit problem with long $PATH
by Tom Terwilliger 25 Jun '09

25 Jun '09

Hi Phenix users, Several of you have had trouble with phenix.autobuild (or phenix.ligandfit) failing if you had a very long $PATH. The problem seems to be that if your $PATH is very long, then adding more to it and re-setting it can lead to a string that is too long (on some systems). Starting with PHENIX version 1.4-100 (today's version) you can add the keyword remove_path_word_list=" cns xiaroot ccp4 phaser" to shorten your $PATH. This keyword will selectively remove all paths from $PATH that include any of the words you specify, and that do not include the word "phenix", when running that phenix job. The log file will list what paths were removed. To figure out what paths to remove, type echo $PATH and choose the ones that identify other programs that are not part of phenix. I hope that will do it! Please let me know if this doesn't solve the problems. All the best, Tom T Thomas C. Terwilliger Mail Stop M888 Los Alamos National Laboratory Los Alamos, NM 87545 Tel: 505-667-0072 email: terwilliger(a)LANL.gov Fax: 505-665-3024 SOLVE web site: http://solve.lanl.gov PHENIX web site: http:www.phenix-online.org ISFI Integrated Center for Structure and Function Innovation web site: http://techcenter.mbi.ucla.edu TB Structural Genomics Consortium web site: http://www.doe-mbi.ucla.edu/TB CBSS Center for Bio-Security Science web site: http://www.lanl.gov/cbss

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