- phenixbb - phenix-online.org

phenix.automr high zscore but too many clashes
by zach powers 17 Apr '09

17 Apr '09

Hi, I have a problem and i wonder if someone has had a similar experience. I have been using phenix.automr program to find an MR solution for a protein. When I ask Phaser to use copies=2 I find a number of solutions with poor z-scores (3-4). When I ask it to use copies=3, I get great Z-scores (see below) but no solution due to the high number of clashes (>100!). As an x-ray newbie I am a bit perplexed of what to make of this: the solution is not good because there is not enough space in the unit cell to comfortably accommodate all three molecules yet the Z-score indicates the structure is good. My structure does have several loops and these may be contributing to the clashing residues. As a newbie I have a newbie question - what does this mean (great z-score but no solutions)? Is this non-solution a possible solution if I play with it, or is the packing simply too tight and the Z-scores are not valid? In the meantime I am chopping my protein into sub-domains as recommended in the Phaser documentation, but if anyone has seen something like this before, any suggestions are welcome. thanks zach charlop-powers Packing Table: Space Group P 3 ------------------------------ Solutions accepted if number of clashes = 135 (lowest number of clashes in list) provided this number of clashes <= 10 (maximum number of allowed clashes) # #Clashes # Accepted Annotation 1 188 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=10.4 2 156 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=10.2 3 151 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=9.8 4 146 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=9.7 5 187 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=16.3 TFZ=10.5 6 202 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=9.2 7 170 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=9.1 8 144 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=16.3 TFZ=10.2 9 177 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=9.1 10 153 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.9 11 150 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.8 12 217 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.6 13 182 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.6 14 135 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.5 15 175 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.3 16 159 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.3 17 192 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.3 18 165 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=16.3 TFZ=9.2 19 169 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=16.3 TFZ=9.1 20 158 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.0 21 173 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=8.0 22 154 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=16.3 TFZ=8.9 23 206 NO RFZ=20.3 TFZ=18.2 PAK=10 LLG=431 RFZ=18.5 TFZ=7.8 0 accepted of 23 solutions

4 4

phenix.autobuild error
by Andreas Förster 15 Apr '09

15 Apr '09

Hey all, I'm calling phenix.autobuild with the following command: phenix.autobuild data=second_try_33.1.mtz model=second_try_33.1.pdb \ > seq_file=name_139_414.seq ps_in_rebuild=True On my MacBook Pro (OSX 5.6, phenix Version: 1.4, Release tag: 3, cctbx tag: 2008_12_07_1353, Platform: mac-intel-osx osx-10.5.6), I get the following error: Exception RuntimeError: 'maximum recursion depth exceeded while calling a Python object' in <type 'exceptions.KeyError'> ignored 108409888 On a linux box (RHEL 5.2, phenix Version: 1.4, Release tag: 3, cctbx tag: 2008_12_07_1353, Platform: intel-linux-2.6 redhat-e5.2) I get this error: ******************************************************************************** Failed to carry out AutoBuild_build_cycle: child process stderr output: command: 'csh /csb/home/andreas/name/P6big/second_try/autobuild/AutoBuild_run_2_/TEMP0/RUN_FILE_1' tset: standard error: Invalid argument ******************************************************************************** Each time, the script was run on the same set of files residing on the linux box. I either ssh into the machine and run phenix.autobuild in an ssh shell or mount my linux home directory via MacFusion and run phenix.autobuild on the Mac. I haven't tried copying the files to the Mac, but other phenix commands work with the remote data (e.g. phenix.autobuild data=data.mtz solvent_fraction=.6 ps_in_rebuild=True model=coords.pdb maps_only=True) What's the problem? Andreas -- Andreas Förster, Research Associate Paul Freemont & Xiaodong Zhang Labs Department of Biochemistry, Imperial College London http://www.msf.bio.ic.ac.uk

2 1

Twin refinement issues?
by Phil Jeffrey 10 Apr '09

10 Apr '09

While experimenting with twin refinement for a crystal in P321 that approximates P622 to varying extents, I noticed: | twin fraction: 0.59 twin operator: -h,-k,l | A bug, perhaps ? This twin fraction would seem to be unlikely, since twin fractions > 0.5 have no meaning. Since this data does not scale well in p622 compared to some more heavily-twinned datasets on this xtal form I think the twin fraction is certainly less than 0.5. Xtriage estimates: Statistics depending on twin laws ----------------------------------------------------------------- | Operator | type | R obs. | Britton alpha | H alpha | ML alpha | ----------------------------------------------------------------- | -h,-k,l | M | 0.150 | 0.343 | 0.347 | 0.283 | ----------------------------------------------------------------- Unless of course phenix.refine is reporting 2*alpha. Phenix v1.4-3 Intel Mac OSX 10.5.6 This particular run with: phenix.refine model-08.pdb pz7e_truncate-unique.mtz refinement.main.ncs=true strategy=individual_sites+group_adp --overwrite xray_data.r_free_flags.generate=True twin_law="-h,-k,l" This is a very early non-finessed model. Phil Jeffrey Princeton

4 6

optimize weighs
by Yusuf Akhter 09 Apr '09

09 Apr '09

Dear all, I am refining a protein structure using phenix.refine, when i tried to use optimize_wxc, optimize_wxu It always crashes and gives following error. May anybody can explain what is the problem?? Traceback (most recent call last): File "/public/xtal/phenix/phenix-1.3b/phenix-1.3b/phenix/phenix/command_line/refine.py", line 5, in <module> command_line.run(command_name="phenix.refine", args=sys.argv[1:]) File "/public/xtal/phenix/phenix-1.3b/phenix-1.3b/phenix/phenix/refinement/command_line.py", line 78, in run call_back_after_monitor_collect=call_back_after_monitor_collect) File "/public/xtal/phenix/phenix-1.3b/phenix-1.3b/phenix/phenix/refinement/driver.py", line 1308, in run log = log) File "/public/xtal/phenix/phenix-1.3b/phenix-1.3b/phenix/phenix/refinement/strategies.py", line 642, in refinement_machine log = log) File "/public/xtal/phenix/phenix-1.3b/phenix-1.3b/phenix/phenix/refinement/weight_xray_chem.py", line 327, in __init__ self.wxc_optimization() File "/public/xtal/phenix/phenix-1.3b/phenix-1.3b/phenix/phenix/refinement/weight_xray_chem.py", line 450, in wxc_optimization rw, rf, b, a, a_xc = self.wx_opt_helper(w = wxc_scale_0 * wxc, sites=True) File "/public/xtal/phenix/phenix-1.3b/phenix-1.3b/phenix/phenix/refinement/weight_xray_chem.py", line 387, in wx_opt_helper verbose = 0) TypeError: __init__() takes at least 5 non-keyword arguments (3 given) Thanks in advance. Yusuf

2 1

twin refinement error
by jonathan elegheert 08 Apr '09

08 Apr '09

Dear Phenix developers, I am rigid-body refining a PM-twinned structure (C2221 => P6x22) with the operators suggested from xtriage or SFcheck but I keep running into this same error (version 1.4.3 or latest cci aps bundles); 1/2*h-1/2*k,-3/2*h-1/2*k,-l 1/2*h+1/2*k,3/2*h-1/2*k,-l Any suggestions as to how I can resolve this? Many thanks in advance, Jonathan Elegheert PhD student Ghent University Belgium ------------------------------------------------------------------------- Traceback (most recent call last): File "/usr/local/cciapps/cci_apps_sources/phenix/phenix/command_line/refine.py", line 16, in <module> command_line.run(command_name="phenix.refine", args=sys.argv[1:]) File "/usr/local/cciapps/cci_apps_sources/phenix/phenix/refinement/command_line.py", line 89, in run call_back_after_monitor_collect=call_back_after_monitor_collect) File "/usr/local/cciapps/cci_apps_sources/phenix/phenix/refinement/driver.py", line 1121, in run call_back_after_monitor_collect = call_back_after_monitor_collect) File "/usr/local/cciapps/cci_apps_sources/phenix/phenix/refinement/strategies.py", line 190, in refinement_machine twin_law = sgtbx.rt_mx( params.twinning.twin_law ) RuntimeError: cctbx InternalError: /net/garter/scratch1/rwgk/auto_build/sources/cctbx/sgtbx/rt_mx.cpp(230): Unsuitable value for rational rotation matrix.

4 6

Phenix.refine Rigid Body multi_body_factor
by Mary Fitzgerald 08 Apr '09

08 Apr '09

I was wondering when you would want to change multi_body_factor= 1? Is this something you would change when you have multiple rigid body domains? From the name, that's what I'm guessing it's for. I'm refining a large over 400 kD multimer complex with two complexes in the ASU. So, if I have 5 chains in each multimer would I want to try to change the multi_body_factor to 10? Or does multi_body_factor affect something else. Thanks, Mary X. Fitzgerald Postdoctoral Associate Arnold Lab

2 1

optimize_wxc and wxu
by Scott Classen 08 Apr '09

08 Apr '09

Hello phenixbb, In general should one optimize wxc and wxu ONLY as a final polishing step or can this be done earlier in the refinement and then the optimized values used for subsequent rounds of refinement? If so which values in the log file should be written into the .def file and where? Here is a snippit from my def file after I optimized weights and changed the optimize flags back to False: target_weights { mode = *automatic every_macro_cycle wxc_scale = 0.5 wxu_scale = 1 wc = 1 wu = 1 fix_wxc = None fix_wxu = None optimize_wxc = False bonds_rmsd_max = 0.05 angles_rmsd_max = 3.5 optimize_wxu = False shake_sites = True shake_adp = 10 regularize_ncycles = 50 verbose = 1 wnc_scale = 0.5 wnu_scale = 1 rmsd_cutoff_for_gradient_filtering = 3 } 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 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

4 3

restrains for nucleaic acid base pairs
by Imre Toeroe 08 Apr '09

08 Apr '09

Hi, Does anybody know an easy way to restrain nucleic acid base pairs (base stacking+Watson-Crick base pairing) in phenix.refine? I am refining against a 3.3 A dataset. Thanks, Imre

3 4

phenix autobuild and omit map
by rpai＠berkeley.edu 07 Apr '09

07 Apr '09

Hi, I have a loop that was improperly built when I ran autobuild. the density according to the Fobs-Fcalc maps is not that great. Someone told me that autobuild can use omit maps to build / rebuild the loops so you dont have to do it by hand. Looking at the manual, I found the command, but am a little confused. The model used is the entire model correct? the target.pdb is the chain or region that was not correctly built. For iterative building...how many cycles should I go for? phenix.autobuild data=w1.sca model=coords.pdb omit_box_pdb=target.pdb \ composite_omit_type=iterative_build_omit thanks Raj

3 2

imaginary map (anomlous difference Fourier) from phenix
by Ulrich Baumann 06 Apr '09

06 Apr '09

Hi everybody, I used phenix for calculating anomalous Fourier coefficients (script below) and I get the desired file containing columns FANOM PHANOM. When I read this into Coot for displaying the imaginary map, I only get noise, regardless of resolution cutoff etc. When I export the Fcalcs as CNS format and use CNS for the map calculation, I get very high peaks (i.e. excellent signal-to-noise ratio) on the anomalous scatterers. Inspection of the phenix fcalc file reveals that FOFCWT_no_fill equals FANOM and PHFOFCWT_no_fill is identical to PHANOM . Obviously I am doing something stupidly wrong. Setting xray_data.force_anomalous_flag_to_be_equal_to=true doesn't change anything, by the way, and i am using the latest Phenix version. I vaguely remember that there were an issue with anomlous data but can't find the thread. Thank you very much in advance for enlightening me, Ulrich # source /usr/local/phenix-1.4-3/phenix_env phenix.refine m226h1_rfree-unique.mtz m226h1_hkl2000_6_waters_001.pdb main.number_of_macro_cycles=1 \ refinement.input.xray_data.labels="F(+),SIGF(+),F(-),SIGF(-)" \ xray_data.high_resolution =3.5 strategy=none export_final_f_model=cns output.prefix=sfcalc --overwrite * Space group = 'P3121' (number 152) The mapcoeff.mtz looks like OVERALL FILE STATISTICS for resolution range 0.000 - 0.319 ======================= Col Sort Min Max Num % Mean Mean Resolution Type Column num order Missing complete abs. Low High label 1 ASC 0 49 0 100.00 23.7 23.7 88.43 1.77 H H 2 NONE 0 28 0 100.00 8.4 8.4 88.43 1.77 H K 3 NONE -69 69 0 100.00 0.6 26.2 88.43 1.77 H L 4 NONE 0.0 2220.8 0 100.00 235.55 235.55 88.43 1.77 F 2FOFCWT 5 NONE -180.0 180.0 0 100.00 2.65 89.99 88.43 1.77 P PH2FOFCWT 6 NONE 0.0 956.4 0 100.00 72.48 72.48 88.43 1.77 F FOFCWT 7 NONE -180.0 180.0 0 100.00 2.16 89.97 88.43 1.77 P PHFOFCWT 8 NONE 0.0 2202.8 117 99.84 234.51 234.51 37.16 1.77 F 2FOFCWT_no_fill 9 NONE -180.0 180.0 117 99.84 2.78 89.99 37.16 1.77 P PH2FOFCWT_no_fill 10 NONE 0.0 961.2 117 99.84 72.39 72.39 37.16 1.77 F FOFCWT_no_fill 11 NONE -180.0 180.0 117 99.84 2.19 89.95 37.16 1.77 P PHFOFCWT_no_fill 12 NONE 0.0 961.2 117 99.84 72.39 72.39 37.16 1.77 F ANOM 13 NONE -180.0 180.0 117 99.84 2.19 89.95 37.16 1.77 P PHANOM No. of reflections used in FILE STATISTICS 72570 1 1 3 88.81 -133.90 400.50 -133.90 139.90 -134.31 502.82 -134.31 502.82 -134.31 1 1 4 719.55 109.38 336.98 109.38 703.51 104.61 304.13 104.61 304.13 104.61 1 1 5 987.30 124.20 664.11 124.20 985.68 119.49 659.96 119.49 659.96 119.49 1 1 6 871.35 35.76 152.79 35.76 894.80 36.84 198.58 36.84 198.58 36.84 1 1 7 425.64 38.73 189.77 38.73 419.88 34.24 177.78 34.24 177.78 34.24 1 1 8 406.00 -154.97 53.51 25.03 416.82 -153.85 32.47 26.15 32.47 26.15 1 1 9 73.73 139.84 441.04 139.84 92.83 139.16 479.45 139.16 479.45 139.16

2 2

maps and autobuild
by chandra bose 06 Apr '09

06 Apr '09

hello, I am a first time user to phenix , and I have just completed the autobuild function,. My question may have been asked many times before, but.. The output after the cycles of autobuild/refinement gave me a refined file for a 2.2 Angstrom data set with an R/Rfree of .28/.32. I want to take a look at the density modified maps and see what I can adjust further, but am having trouble loading it in coot. Am I correct in understanding that Coot can load and construct 2f0-fc maps? if so How? : exptl_fobs_phases_freeR_flags.mtz overall_best_denmod_map_coeffs.mtz; overall_best_refine_map_coeffs.mtz; Also from reading the manual, as I understand it, the phenix.refine command will automatically generate Coot compatible maps. But if I take my autobuild.pdb file and the original mtz data file, how can I quickly obtain the same R/Rfree, what commands would I have to imput other than phenix.refine mtz pbd? Also, is it possible to make 3Fo-2Fc maps? Thanks Charlie

3 2

phenix.autobuild sa_omit map
by Jianghai Zhu 03 Apr '09

03 Apr '09

Hi, I am using phenix.autobuild to calculate a sa_omit map. The find_ncs is Auto by default. What does Auto mean here? Should I turn it off or does it matter? Thanks. -- Jianghai

2 1

refinment of ligand
by peter hudson 01 Apr '09

01 Apr '09

Hello all I have refined and built model a dataset of 3.0A resolution dataset. This model is assocaied with a ligand. After final refinement and model building i found a big blob of Fo-Fc density of around 4sigma level at the N-terminal of the fianl model. My protein doesnot carry any tag at the N-terminal. But, the template i have used for the molecular replacement carrying a tag at the N-termianl, which can occupy only 1/4th of this positive density after superpostion. My crystallisation conditions component cannot occupy such higher level of positive density. Since my protein binds to a ligand, i looked carefully to the positive density and it seems very similar to the ligand density but its obscure. Refinement after fitting the ligand leads to very high B-factor of the ligan, which vary for various atoms from 50-90 and simultaneously positive density goes off. I also tried to change the occupancy of the ligand but as i reduce the occupancy the B factor comes down at the noraml expected average B factor value but again Fo-Fc density appears in the map over ligand. If i leave to refine the occupancy to the programme automatically, this lead to the zero occupancy of the few atoms in the ligand and avearge B factor remains normal. suggestions would be appreciated. Thanks in advance Peter

1 0

phenix.autobuild disappeared
by Andreas Förster 01 Apr '09

01 Apr '09

Hey there, sorry for the brainfreeze but I can't find phenix.autobuild. It's not part of cci_apps Versions 2008_09_13_0905 or 2009_02_15_2320. What happened? How do I build? Andreas -- Andreas Förster, Research Associate Paul Freemont & Xiaodong Zhang Labs Department of Biochemistry, Imperial College London http://www.msf.bio.ic.ac.uk

2 1

Re: [phenixbb] phenixbb Digest, Vol 40, Issue 31
by Marjolein Thunnissen 31 Mar '09

31 Mar '09

> > Hi Marjolein, > >> I have been trying to find in the documentation if there is a way to do this >> but I have the following problem: In a recent structure we are currently >> refining we identified a spermine molecule sitting right on a >> crystallographic two-fold axis. Spermine has internal two-fold symmetry and >> it is part of extensive crystal contacts. I just wonder how to refine this >> with Phenix. In Refmac it is easy, I put the occupancies to 0.5 and the >> program ignores the clashes of the symmetry related molecule, but that >> doesn't seem to be the way that Phenix works. Can anyone give me advise on >> this? > > Supporting the occupancy trick is on my to-do list (fairly high up). > > In the meantime, could you try giving only the asymmetric unit of the > spermine? I.e. delete all duplicate atoms from your pdb file. > The restraints across the symmetry bond will not work automatically. > You have to define a "custom bond" instead, and if you also want to > restrain the angles, custom pseudo-bonds. See the > "Definition of custom bonds and angles" section. > In your case you'll have to define symmetry_operations (not shown > in the docs), e.g.: > > bond { > action = *add > atom_selection_1 = resname SPM and name C7 > atom_selection_2 = resname SPM and name C7 > symmetry_operation = -x,-y,z > distance_ideal = 1.5 > sigma = 0.01 > } > > The main difficulty will be to find the correct symmetry_operation > for each custom bond. If you send my the CRYST1 card from your > PDB file and the part with the SPM residue I could help out. > > Ralf > > > ------------------------------ Hi, The commands worked indeed the trick. Luckily it is actually not that difficult to find the symmetry operators when using coot. One just has to switch drawing symmetry atoms on and clicking on a symmetry atom, coot will tell you in the bottom line which atom it is and which symmetry operator was used to draw it... Thanks again Marjolein __________________________________________________________________________ | Marjolein Thunnissen Phone +46-(0)46-22 24584| | Associate Professor Fax +46-(0)46-22 24692| | Dept of Molecular Biophysics, Lund University http://www.mbfys.lu.se | | PO-Box 124 S-221 00 Lund, Sweden | | | |Scientific coordinator I911 (Max-lab): MAD and fixed-wavelength stations | | for macromolecular crystallography | |__________________________________________________________________________|

2 1

Re: [phenixbb] PHENIX user mailing list <[email protected]>
by Ralf W. Grosse-Kunstleve 30 Mar '09

30 Mar '09

Hi Maia, > I have a Ca2+ ion in a special position. the ocupancy should be 0.5, as > the other half belongs to a symmetry-related molecule. However, > refinement makes the occupansy 1. What should I do to prevent this? > Should not it find automatically that? phenix.refine uses the correct symmetry factor automatically (internally). Unless you have an actual partial occupancy, the occupancy factor in the pdb file should be 1, irrespective of the multiplicity of the special position. Let me know if this doesn't answer your question. Ralf

4 4

can I turn off TLS for subsequent rounds of refinement?
by Scott Classen 30 Mar '09

30 Mar '09

Hi all, As the TLS refinement can take a significant amount of time, I'm wondering if I can turn it off after a few rounds with it on? or will phenix.refine "see" the ANISOU records and assume I am doing anisotropic ADP? Thanks, Scott

2 1

phenix automr
by sbiswas2＠ncsu.edu 30 Mar '09

30 Mar '09

Hi all, I tried to use phenix automr by using the gui and also I am using the latest version of phenix (courtesy of the phenix development team). I asked it to search for two molecules in the asymmetric unit but it finds just one also I am getting negative (-29) LLG value for the final solution. The map also does not look good. I also asked it to search for all possible space group, although I processed the data with P43212 phenix automr finds a different one P41212.Will fixing the space group fix the problem? Does anyone know what is going on? I tried running phaser separately through ccp4 and phaser did find two molecules although the map did not look good in this either. Shya

5 4

Missing map for model
by Folmer Fredslund 30 Mar '09

30 Mar '09

Dear all, In phenix.refine (1.4-3) my electron density maps look weird. I get beautiful density for a helix that is missing from the current model, but absolutely horrible density for the rest of the molecule. Any ideas what is going on? Maps from automr look fine and the density modified map from autobuild also look fine, whereas the autobuild map from refinement has the same features. Any ideas what is going on? I am running all programs with default parameters. Best regards, Folmer Fredslund

2 3

Geometry problem with the ligand
by Jhon Thomas 29 Mar '09

29 Mar '09

Hello all I am working with a 3.0 A dataset, which binds with cAMP. I have refined the dataset with phenix (TLS+Group_adp) and followed by weight optimization. Though cAMP fit well in the density, validation tools (molprobity, procheck) suggest the bond angles and bond lengths of ligand of completely refined model is very bad(100% bad). Can anyone suggest what could be the problem with model or refinement? I will appreciate the suggestions. Thanks in advance John

3 2

weight optimization
by peter hudson 29 Mar '09

29 Mar '09

Hello all I am very new to this field. I am refining a dataset of 3.0A resolution. I want to perform the weight optimization. while weight optimisation, occupancy refinemnet is default. I donot want torefine the occupancy of any of the atoms from my model. How can i set to occupancy_refinemnet=false. I will appreciate the suggestions. Thanks in advance Peter

3 2

Re: [phenixbb] Question about scale of ED maps from phenix vs map coeffs when displayed in COOT
by Ralf W. Grosse-Kunstleve 28 Mar '09

28 Mar '09

For phenix maps: the sum is over the entire unit cell. Ralf > Is the sum over the asymmetric unit or the portion of space that > happened to be covered by the map? The result of this calculation > depends greatly on the amount of solvent region that is included in > the sum. > > Dale Tronrud > > Ralf W. Grosse-Kunstleve wrote: > >> That was (and is) always a mystery for me: when I look at two maps (both > >> coming from phenix.refine): > >> - one is computed in COOT from phenix.refine map coefficients, and > >> - another one displayed in PyMol using X-plor formatted map. > >> > >> To make both maps looking identical I have to draw them at different > >> sigma levels, sometime very different. This is true for both fo-fc and > >> 2fo-fc maps. > >> > >> I have some ideas but no any proofs so I don't want to put my > >> speculations to the bb. So, I have no answer... May be Ralf or Paul E...? > > > > Our "sigmas scaled maps" are simply divided by the "biased standard > > deviation": > > > > sqrt(sum((value-mean)**2) / n) > > > > The mean of our maps is usually zero, as we don't usually include F000. > > > > Ralf

3 2

Refinement error
by Katya Heldwein 27 Mar '09

27 Mar '09

Hi, I am getting the following error when I run phenix.refine. How can I get rid of it? Katya =============================== refinement start ============================== ----------structure factors based statistics (before refinement)---------- ----------X-ray data---------- show_stack(1): /private/var/automount/nfs/programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/mmtbx/twinning/twin_f_model.py(1676) r_values show_stack(2): /private/var/automount/nfs/programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/mmtbx/twinning/twin_f_model.py(1837) r_work show_stack(3): /private/var/automount/nfs/programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/mmtbx/f_model.py(2249) __init__ show_stack(4): /private/var/automount/nfs/programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/mmtbx/twinning/twin_f_model.py(1139) info show_stack(5): /private/var/automount/nfs/programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/mmtbx/__init__.py(109) show_comprehensive show_stack(6): /private/var/automount/nfs/programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/phenix/phenix/refinement/strategies.py(280) refinement_machine show_stack(7): /private/var/automount/nfs/programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/phenix/phenix/refinement/driver.py(1121) run show_stack(8): /private/var/automount/nfs/programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/phenix/phenix/refinement/command_line.py(89) run show_stack(9): /programs/i386-mac/cciapps/2009_02_15_2320/cci_apps_sources/phenix/phenix/command_line/refine.py(16) <module> Floating-point error (Python call stack above) This crash may be due to a problem in any imported Python module, including modules which are not part of the cctbx project. To disable the traps leading to this message, define these environment variables (e.g. assign the value 1): BOOST_ADAPTBX_FPE_DEFAULT BOOST_ADAPTBX_SIGNALS_DEFAULT This will NOT solve the problem, just mask it, but may allow you to proceed in case it is not critical. -- Ekaterina Heldwein, Ph.D. Assistant Professor Department of Molecular Biology and Microbiology Tufts University 136 Harrison Ave Boston, MA 02111 phone: 617-636-0858 fax: 617-636-0337 e-mail: katya.heldwein(a)tufts.edu

4 3

AssertionError in twin refinement...
by Francis E Reyes 26 Mar '09

26 Mar '09

====================== bulk solvent modeling and scaling ====================== |--(resolution: 2.00 - 23.08 A; n_refl. = 24304)------------------------------| | | | r_work= 0.2462 r_free= 0.2706 ksol= 0.32 Bsol= 47.55 scale= 1.149 | | | | overall anisotropic scale matrix (Cartesian basis; B11,B22,B33,B12,B13,B23):| | (-2.11,5.80,-3.69,0.00,-0.00,-0.00); trace/3= -0.00 | | | | maximum likelihood estimate for coordinate error: None A | | x-ray target function (twin_lsq_f) for work reflections: 0.346916 | | twin fraction: 0.03 twin operator: k,h,- l | | -----------------------------------------------------------------------------| ================== ordered solvent: location/analysis/update ================== Start model: number = 165 b_iso_min = 37.62 (limit = 1.00) b_iso_max = 70.26 (limit = 50.00) b_iso_mean = 49.33 anisotropy_min = 1.00 (limit = 0.10) occupancy_min = 1.00 (limit = 1.00) occupancy_max = 1.00 (limit = 1.00) dist_sol_mol_min = 2.45 (limit = 1.80) dist_sol_mol_max = 4.94 (limit = 6.00) Filtered: number = 71 b_iso_min = 37.71 (limit = 1.00) b_iso_max = 49.89 (limit = 50.00) b_iso_mean = 45.14 anisotropy_min = 1.00 (limit = 0.10) occupancy_min = 1.00 (limit = 1.00) occupancy_max = 1.00 (limit = 1.00) dist_sol_mol_min = 2.45 (limit = 1.80) dist_sol_mol_max = 3.99 (limit = 6.00) Traceback (most recent call last): File "/usr/local/phenix-1.3-final/phenix/phenix/command_line/ refine.py", line 5, in <module> command_line.run(command_name="phenix.refine", args=sys.argv[1:]) File "/usr/local/phenix-1.3-final/phenix/phenix/refinement/ command_line.py", line 89, in run call_back_after_monitor_collect=call_back_after_monitor_collect) File "/usr/local/phenix-1.3-final/phenix/phenix/refinement/ driver.py", line 1081, in run call_back_after_monitor_collect = call_back_after_monitor_collect) File "/usr/local/phenix-1.3-final/phenix/phenix/refinement/ strategies.py", line 364, in refinement_machine log = log) File "/usr/local/phenix-1.3-final/mmtbx/mmtbx/solvent/ ordered_solvent.py", line 159, in __init__ use_kick_map = self.params.use_kick_maps).peaks_mapped() File "/usr/local/phenix-1.3-final/mmtbx/mmtbx/solvent/ ordered_solvent.py", line 332, in find_peaks log = self.log) File "/usr/local/phenix-1.3-final/mmtbx/mmtbx/find_peaks.py", line 85, in __init__ symmetry_flags = maptbx.use_space_group_symmetry) File "/usr/local/phenix-1.3-final/mmtbx/mmtbx/twinning/ twin_f_model.py", line 2244, in electron_density_map "m_gradient") AssertionError phenix version is 1.3 final. Ideas? thanks! FR --------------------------------------------- Francis Reyes M.Sc. 215 UCB University of Colorado at Boulder gpg --keyserver pgp.mit.edu --recv-keys 67BA8D5D 8AE2 F2F4 90F7 9640 28BC 686F 78FD 6669 67BA 8D5D

2 7

Spermine molecule on a crystallographic two-fold axis
by Marjolein Thunnissen 26 Mar '09

26 Mar '09

I have been trying to find in the documentation if there is a way to do this but I have the following problem: In a recent structure we are currently refining we identified a spermine molecule sitting right on a crystallographic two-fold axis. Spermine has internal two-fold symmetry and it is part of extensive crystal contacts. I just wonder how to refine this with Phenix. In Refmac it is easy, I put the occupancies to 0.5 and the program ignores the clashes of the symmetry related molecule, but that doesn't seem to be the way that Phenix works. Can anyone give me advise on this? Thanks __________________________________________________________________________ | Marjolein Thunnissen Phone +46-(0)46-22 24584| | Associate Professor Fax +46-(0)46-22 24692| | Dept of Molecular Biophysics, Lund University http://www.mbfys.lu.se | | PO-Box 124 S-221 00 Lund, Sweden | | | |Scientific Coordinator I911 (Max-lab): MAD and fixed-wavelength stations | | for macromolecular crystallography | |__________________________________________________________________________|

2 1

twin refine error message
by Alex Taylor 26 Mar '09

26 Mar '09

Hi, I am trying to refine using the following twin operators output from xtriage for C2221 1/2*h-1/2*k,-3/2*h-1/2*k,-l 1/2*h+1/2*k,3/2*h-1/2*k,-l but phenix.refine (vers 1.4-4 and 1.4-3) fails with the following error message: =============================== refinement start ============================== Traceback (most recent call last): File "/usr/progs/linux/phenix-1.4-4/phenix/phenix/command_line/refine.py", line 18, in <module> command_line.run(command_name="phenix.refine", args=sys.argv[1:]) File "/usr/progs/linux/phenix-1.4-4/phenix/phenix/refinement/command_line.py", line 89, in run call_back_after_monitor_collect=call_back_after_monitor_collect) File "/usr/progs/linux/phenix-1.4-4/phenix/phenix/refinement/driver.py", line 1121, in run call_back_after_monitor_collect = call_back_after_monitor_collect) File "/usr/progs/linux/phenix-1.4-4/phenix/phenix/refinement/strategies.py", line 191, in refinement_machine twin_law = sgtbx.rt_mx( params.twinning.twin_law ) RuntimeError: cctbx InternalError: /net/anaconda/scratch1/phenix/phenix-1.4-4/cctbx_project/cctbx/sgtbx/rt_mx.cpp(230): Unsuitable value for rational rotation matrix. I am running the RedHat WS 5.2 64-bit OS. Thanks for suggestions, Alex -- Alex Taylor, Ph.D. Technical Director, X-ray Crystallography Core Laboratory University of Texas Health Science Center at San Antonio Office: 210-567-3781 FAX: 210-567-6595

2 1

Postdoc Crystallographer Position - Berkeley
by Paul Adams 24 Mar '09

24 Mar '09

Lawrence Berkeley Laboratory Postdoctoral Researcher Job ID: 22539 Division: Physical Biosciences Date Opened: 3/24/2009 Summary: A postdoctoral position is immediately available to study structure and function of macromolecular complexes and enzymes using X-ray crystallographic methods. This position is joint between the Center for Protein Folding Machinery (www.proteinfoldingcenter.org) and the Joint BioEnergy Institute (www.jbei.org). Robotic hardware for performing crystallization trials and imaging results are available. Crystals will be characterized and data collected using the beamline resources of the Berkeley Center for Structural Biology at the Advanced Light Source. The biological systems to be studied are: - Type-II chaperonins responsible for the refolding of unfolded proteins in archeal and mammalian cells. - Glycosyl transferases responsible for the synthesis of hemicellulose in plants. - Novel glycosyl hydrolases that are able to breakdown cellulosic material for biofuels production. Duties: Using robotic hardware to perform screens for crystallization conditions. Optimization of crystallization conditions. Biophysical characterization of protein samples using standard techniques, such as dynamic light scattering. Characterization of crystals using the Berkeley Center for Structural Biology beamlines at the Advanced Light Source. Collection and analysis of diffraction data, model building and structure refinement. Extensive email and verbal interaction with other researchers. Qualifications: Ph.D. or equivalent in a scientific discipline, preferably structural biology, biology, or chemistry. Demonstrated experience of scientific research using crystallographic methods. Experience with crystallization methods. Solid interpersonal skills and the ability to work in a team environment are critical. Ability to communicate with a broad range of researchers. Note that this is a one-year appointment with renewal based on performance and continued funding. Lawrence Berkeley National Laboratory is a world leader in science and engineering research, with 11 Nobel Prize recipients over the past 75 years, and 59 present members of the National Academy of Sciences. LBNL conducts unclassified research across a wide range of scientific disciplines and hosts four national user facilities. AA/EEO employer committed to the development of a safe and diverse workforce. Learn more at http://www.lbl.gov. To apply visit http://cjo.lbl.gov/ and search for the job number 22539 or visit: http://jobs.lbl.gov/LBNLCareers/details.asp?jid=22539&p=1 -- Paul Adams Deputy Division Director, Physical Biosciences Division, Lawrence Berkeley Lab Adjunct Professor, Department of Bioengineering, U.C. Berkeley Vice President for Technology, the Joint BioEnergy Institute Head, Berkeley Center for Structural Biology 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: Patty Jimenez [ PAJimenez(a)lbl.gov ] [ 1-510-486-7963 ] --

1 0

refinement with a covalent inhibitor.
by Maia Cherney 23 Mar '09

23 Mar '09

Hi all, I am trying to refine a complex with a covalent inhibitor (E-64) that has a carboxylate group. My model has it planar, but after refinement the two oxygens of the carboxyl group get out of the plane. In the cif file generated by elbow this plane description is missing, I added it manually, but the result is the same. Maia

4 11

ADP settings for protein and solvent
by Katya Heldwein 23 Mar '09

23 Mar '09

Hi, I'd like to refine group adps for my protein and individual adps for the solvent while refining coordinates for all atoms. How can I set that up in the command line mode? Katya -- Ekaterina Heldwein, Ph.D. Assistant Professor Department of Molecular Biology and Microbiology Tufts University School of Medicine 136 Harrison Ave Boston, MA 02111 phone: 617-636-0858 fax: 617-636-0337 e-mail: katya.heldwein(a)tufts.edu

2 1

Question about scale of ED maps from phenix vs map coeffs when displayed in COOT
by Schubert, Carsten [PRDUS] 20 Mar '09

20 Mar '09

Hi, Sorry for the crosspost, but I am not sure were the problem is located: I noticed that the xplor maps generated by phenix seem to be on a different scale than the maps generated from the map-coefficients from the same refinement run when the maps-coeffs are read into coot. Usually I only read in the map-coeffs and the scale seems reasonable to me, i.e. average density ~0.28 e/A3 at 1 sigma for a 2fofc map. The map from the same run (converted to ccp4 map in mapman) is at 0.55e/A3 at 1 sigma, when displayed in coot. The map should be normalized in phenix (apply_sigma_scaling=True) and the stats from mapman indicate an RMS deviation of ~1. For fofc maps I need to crank up the display to 6 sigma to get an equivalently looking map as the one from the map-coefficients at 3 sigma contour level. Any hints as to were the factor of 2 comes from, coot or phenix? Cheers, Carsten

3 2

Problems with phasing a protein (1300aa)
by Kumar 20 Mar '09

20 Mar '09

Hello Phenixbb members, I have been trying to obtain phases for a protein which contain ~1300aa. We have obtained native data to a resolution of 3.3A (Space group I222 or I212121). But we are having tough time phasing it. 'Se' labeled crystals diffracts maximally up to 3.5 to 4 A and dies very quickly on most of the beamlines. We have scanned at Se wavelength and it gives very strong signal as it contain ~45 Se in AU (1300 aa). It is difficult to collect a complete dataset (maximally we get 50-60 % completion with Rmerge ~15) out of one crystal on regular beamline. At microfocus beamline (APS), we were able to collect data in 3-4 batches and merge them to get a complete dataset (Rmerge ~18-20) out of one crystal. We used data collected on microfocus beamline (at peak wavelength) for locating heavy atom position using SHELXD, Solve and Phenix.hyss. SOlve and Phenix.hyss find very few heavy atom sites 1-5 whereas SHELX-CDE lists many but shows no difference in original and inverted (contrast and connectivity). Our phasing attempts with datasets obtained after merging two incomplete dataset from two different crystal has also been disappointing. My another worry is absolute value of average intensity, which seems to be quite low in most of the datasets. Below I have pasted last table of scale.log (HKL2000). Shell Lower Upper Average Average Norm. Linear Square limit Angstrom I error stat. Chi**2 R-fac R-fac 50.00 7.53 45.4 1.6 1.3 1.295 0.055 0.047 7.53 5.98 11.4 1.3 1.3 0.672 0.135 0.114 5.98 5.23 11.2 1.6 1.6 0.643 0.171 0.152 5.23 4.75 16.8 2.0 1.9 0.736 0.148 0.118 4.75 4.41 18.8 2.2 2.2 0.739 0.143 0.132 4.41 4.15 14.6 2.4 2.4 0.653 0.190 0.175 4.15 3.94 11.3 2.5 2.5 0.582 0.247 0.226 3.94 3.77 10.1 2.8 2.8 0.511 0.280 0.191 3.77 3.63 8.0 3.1 3.1 0.450 0.315 0.285 3.63 3.50 7.6 3.3 3.2 0.483 0.311 0.270 All reflections 15.5 2.3 2.2 0.694 0.153 0.106 Now, I want you to help me by answering some of my queries: 1. Is it possible to get MAD/SAD phasing done from a dataset having more than 15% Rmerge and resolution in the range of 4 - 4.5 Ang? 2. Will a complete data set obtained from merging various batches(30-40 frames each) from one or more than one crystal will have proper anomalous signal for phasing? I am worried as weak anomalous signal may get lost while merging. 3. Will such a low value of average Intensities (as shown above from HKL scale log file) will be good enough for MAD/SAD phasing or I really need to improve crystal quality for stronger diffraction. 4. For MAD/SAD phasing, till what resolution we need to have anomalous signal ? Many of my datasets shows anomalous signal maximally up to 6-8 A (calculated using Phenix.xtriage). 5. Since I have low resolution (3.5 to 4 A)data, relatively high Rmerge (14-15%), lower value of average intensity, anomalous signal up to 6 A or so..... which programs will be more useful for heavy atom location and to prevent false positives from being selected? We have been also trying our luck with heavy atom soak but that also has not been very encouraging. I would appreciate any suggestions in this regard. Thanks in advance and sorry for such a long mail. Kumar

4 4

question about generating cif and pdb file using elbow
by crystallogrphy 20 Mar '09

20 Mar '09

Hi, I am using phenix.elbow to generate pdb and cif files with the tripos format file: phenix.elbow --initial_geometry egcg.mol2 --opt --output="LIG" --pdb-ligand The input pdb file contains hydrogen atoms and the output pdb file also contains hydrogen atom. My crystal data only difftracted to 2.4A, so hydrogen atoms cannot be seen. In the refinement, my ligand should not contain hydrogen atoms. So I add one more option not to add hydrogen following the online phenix documentation: phenix.elbow --initial_geometry --write-hydrogens=False egcg.mol2 --opt --output="LIG" --pdb-ligand *However, the program failed. Does anyone know why?* Usage: phenix.elbow [options] *phenix.elbow: error: no such option: --write-hydrogens* -- 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

6 5

Problems with kicked maps in 1.4.3 or 1.4.4
by Schubert, Carsten [PRDUS] 20 Mar '09

20 Mar '09

Hi Pavel, I am having some problems with kicked maps. The job fails with a cryptic message: "Sorry Duplicate mtz_label_amplitudes: 2FOFCWT" and nothing gets written I noticed this in 1.4.3 a couple of days ago, just upgraded to 1.4.4, same problem. Running the same job w/o kicked maps enabled runs fine. Any ideas? Cheers, Carsten Here is a snipped of the .def file. output { prefix = "Prim_105_D6" serial = 4 serial_format = "%03d" write_eff_file = True write_geo_file = False write_def_file = True export_final_f_model = mtz cns write_maps = True write_map_coefficients = True } electron_density_maps { map_format = *xplor map_coefficients_format = *mtz phs suppress = None map { mtz_label_amplitudes = None mtz_label_phases = None likelihood_weighted = None obs_factor = None calc_factor = None kicked = False fill_missing_f_obs_with_weighted_f_model = True } map { mtz_label_amplitudes = "2FOFCWT_no_fill" mtz_label_phases = "PH2FOFCWT_no_fill" likelihood_weighted = True obs_factor = 2 calc_factor = 1 kicked = False fill_missing_f_obs_with_weighted_f_model = False } map { mtz_label_amplitudes = "FOFCWT_no_fill" mtz_label_phases = "PHFOFCWT_no_fill" likelihood_weighted = True obs_factor = 1 calc_factor = 1 kicked = False fill_missing_f_obs_with_weighted_f_model = False } map { mtz_label_amplitudes = "2FOFCWT" mtz_label_phases = "PH2FOFCWT" likelihood_weighted = True obs_factor = 2 calc_factor = 1 kicked = True fill_missing_f_obs_with_weighted_f_model = True } map { mtz_label_amplitudes = "FOFCWT" mtz_label_phases = "PHFOFCWT" likelihood_weighted = True obs_factor = 1 calc_factor = 1 kicked = True fill_missing_f_obs_with_weighted_f_model = True } anomalous_difference_map { mtz_label_amplitudes = "ANOM" mtz_label_phases = "PHANOM" } grid_resolution_factor = 0.25 region = *selection cell atom_selection = None atom_selection_buffer = 3 apply_sigma_scaling = True apply_volume_scaling = False }

2 3

Re: [phenixbb] Can Phenix do MR with medium/low res EM models?
by Gino Cingolani 19 Mar '09

19 Mar '09

thanks Ed, I'll look it up.... I kind of tried something similar but never worked in my hands. I'll give it another try! By the way, the address of my house is 107 Ardsley Drive Dewitt 13214, NY. See you on Saturday 10 or so. Give me a call if you have problems 315 657 4805 Gino *********************************************************** Gino Cingolani, Ph.D. Assistant Professor Dept. of Biochemistry and Molecular Biology SUNY Upstate Medical University 750 E. Adams Street, Syracuse, NY, 13210 Tel. (315) 464 8744 Fax. (315) 464 8750 Email: cingolag(a)upstate.edu *********************************************************** "Nati non foste per viver come bruti, ma per seguir virtute e conoscenza" ("You were not born to live like brutes, but to follow virtue and knowledge") Dante, The Divine Comedy (Inferno, XXVI, vv. 119-120)

2 1

IndexError
by Huiying Li 19 Mar '09

19 Mar '09

I was running phenix refinement using the following command: phenix.refine data.mtz model.pdb \ strategy=individual_sites+tls+individual_adp \ my_lib.cif tls.params the run crashed with the error: ...... ...... File "/usr/local/phenix-1.3-final/mmtbx/mmtbx/monomer_library/pdb_interpretation.py", line 970, in get_lib_link atom_dicts[angle.atom_3_comp_id].get(angle.atom_id_3, None)] IndexError: list index out of range Is the index error refering to some problem in the reflection file (mtz)? Thanks for any suggestions. Huiying _____ Huiying Li, Ph. D Department of Molecular Biology and Biochemistry Natural Sciences I, Rm 2443 University of California at Irvine Irvine, CA 92697, USA Tel: 949-824-4322(or -1953); Fax: 949-824-3280 email: hli(a)uci.edu

2 1

Can Phenix do MR with medium/low res EM models?
by Gino Cingolani 19 Mar '09

19 Mar '09

Hi Phenix community, I was wondering if Phenix can handle MR starting from medium/low res EM models. I have a very large oligomeric protein (2.4MDa) with lots of NCS and >450 Se-Mets. In principle, I could use low res MR phases from the "replaced" model to locate Se-sites and then NCS-averaging to locate more sites and improve my phases. Any suggestions how to get Phenix to use an EM model? Has anybody done it successfully using Phenix? Thanks in advance, Gino *********************************************************** Gino Cingolani, Ph.D. Assistant Professor Dept. of Biochemistry and Molecular Biology SUNY Upstate Medical University 750 E. Adams Street, Syracuse, NY, 13210 Tel. (315) 464 8744 Fax. (315) 464 8750 Email: cingolag(a)upstate.edu *********************************************************** "Nati non foste per viver come bruti, ma per seguir virtute e conoscenza" ("You were not born to live like brutes, but to follow virtue and knowledge") Dante, The Divine Comedy (Inferno, XXVI, vv. 119-120)

2 1

Re: [phenixbb] Can Phenix do MR with medium/low res EM models?
by Thomas C. Terwilliger 19 Mar '09

19 Mar '09

Hi Gino, I have never tried any problem of this size...but here is a way to proceed using Randy's "input_partpdb_file" option for Phaser that is now available in the autosol wizard. Take your EM model with all its NCS copies, and give it to AutoSol with the keywords: input_partpdb_file=MR.pdb partpdb_rms= 3.0 The partpdb_rms should be the estimated RMSD between the MR model and the true model. You will have as good an idea about this as anyone... You also would supply the usual information to autosol: phenix.autosol sad-data.sca seq.dat 450 Se f_prime=-8 f_double_prime=4.5 \ build=False input_partpdb_file=MR.pdb\ partpdb_rms=3.0 \ resolution=6.0 You might cut the resolution to a pretty low value like 6 A as above. If you get a lot of good sites refined by phaser, then you may be on the right track. This autosol run will output phaser_1.mtz and also resolve_1.mtz that you can use as a starting point for further density modification and model-building as appropriate. Given the size of this problem, I would anticipate lots of challenges, and we will be happy to help as you carry on with it. You might try running everything at really low resolution (20A) at first just to get things to run... All the best, Tom T Hi Phenix community, I was wondering if Phenix can handle MR starting from medium/low res EM models. I have a very large oligomeric protein (2.4MDa) with lots of NCS and >450 Se-Mets. In principle, I could use low res MR phases from the "replaced" model to locate Se-sites and then NCS-averaging to locate more sites and improve my phases. Any suggestions how to get Phenix to use an EM model? Has anybody done it successfully using Phenix? Thanks in advance, Gino *********************************************************** Gino Cingolani, Ph.D. Assistant Professor Dept. of Biochemistry and Molecular Biology SUNY Upstate Medical University 750 E. Adams Street, Syracuse, NY, 13210 Tel. (315) 464 8744 Fax. (315) 464 8750 Email: cingolag(a)upstate.edu *********************************************************** "Nati non foste per viver come bruti, ma per seguir virtute e conoscenza" ("You were not born to live like brutes, but to follow virtue and knowledge") Dante, The Divine Comedy (Inferno, XXVI, vv. 119-120) _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org http://www.phenix-online.org/mailman/listinfo/phenixbb

1 0

omit maps without density modification?
by NATHAN THOMSEN 17 Mar '09

17 Mar '09

Hi, Is there is a way to calculate simple omit maps and sa-omit maps in phenix without using density modification? The density modification seems to be flattening the weaker regions of my map - when compared to omit maps from cns for instance. Thanks! Nathan

2 1

AutoBuild error: csh word too long
by Zheng Zhou 16 Mar '09

16 Mar '09

Hi, All I am new to phenix. I met the following error when I run autobuild. I also found the following link from Dr. Lang asking the similar question. But I didn't find a clear solution. Thanks for your help. http://www.phenix-online.org/pipermail/phenixbb/2008-June/001087.html Starting job 1... ******************************************************************************** Failed to carry out AutoBuild_build_cycle: child process stderr output: command: 'csh /home/zz/AutoBuild_run_1_/TEMP0/RUN_FILE_1' Word too long. ******************************************************************************** Zheng

2 1

Covalent ligands
by cdas＠purdue.edu 14 Mar '09

14 Mar '09

Greetings everyone: I have been dealing with a structure of a protein that has a Cys residue covalently modified with a peptide fluromethyl ketone. Can someone tell me how to include this ligand in phenix.refine? In the past, I have used CNS for this purpose. I created a topology and parameter file (using ProDrug server) and modified a patch in the refine script in CNS to establish a covalent connection between an atom in the ligand and gamma sulfur of the protein in question. Thanks. Chitta

4 3

Beamtime proposal deadline for ALS GU time
by Peter Zwart 12 Mar '09

12 Mar '09

The deadline for the may/June cycle of general user time for PX in the ALS is coming up soon (This Sunday). The Berkeley Center for Structural Biology (BCSB) has recently completed a number of upgrades: Sector 8: Recent improvements include: * A high-accuracy microdiffractometer on BL 8.2.1 allows for centering and viewing of very small crystals; * Robot automounters on BL8.2.1 and 8.2.2 are now fully commissioned and the beamlines can be used remotely; * A new large format (315mm active surface area) CCD detector on BL 8.2.1 facilitates data collection on large unit cell crystals. Sector 5: Recent improvements include: * Beamlines (5.0.1 and 5.0.3) now generate 1.5x1011 photons per second at 0.97 Angstrom. This wavelength is slightly above the Se K edge making them ideal for performing Se SAD experiments. * Beamline 5.0.2 (the tunable beamline) generates a peak flux of 8x1011 photons per second, and the energy range of 5.0.2 has been extended to 17 keV, enabling the routine use of shorter wavelengths and anomalously scattering elements such as bromine. * All sector 5 beamlines can be run remotely. The automated sample mounter holds six pucks (96 sample in total). On weekdays, staff is available 22 hours (out of 24) for puck changes and assistance. Please visit http://bcsb.lbl.gov/ for more details about the Center and its beamlines. If you'd like to apply for May/June beamtime at the Advanced Light Source, please submit a General User proposal by March 15, 2009. To find out more, click on: http://www-als.lbl.gov/als/quickguide/independinvest.html We invite you to submit a proposal at: http://alsusweb.lbl.gov/4DCGI/WEB_GetForm/PXProposalEntry.shtml/Initialize. If you have any questions or would like to request open beamtime, please e-mail BCSBBeamtime(a)lbl.gov. [Please note that executed user agreements must be received by LBNL prior to beamtime. Proprietary fees, if applicable, must be received by LBNL at least five working days prior to scheduled beamtime.] -- ----------------------------------------------------------------- P.H. Zwart Beamline Scientist Berkeley Center for Structural Biology Lawrence Berkeley National Laboratories 1 Cyclotron Road, Berkeley, CA-94703, USA Cell: 510 289 9246 BCSB: http://bcsb.als.lbl.gov PHENIX: http://www.phenix-online.org CCTBX: http://cctbx.sf.net -----------------------------------------------------------------

1 0

Re: [phenixbb] phenix.multi_crystal_average with twinned data
by Thomas C. Terwilliger 11 Mar '09

11 Mar '09

Hi Peter and Mary, For the cross-crystal averaging procedure, no refinement is done, so no problem about using detwinned data in phenix.refine. in this case. If detwinned data is supplied then it will be used for density modification. That is probably the best procedure at present. (Note that when running autobuild, you can input a specific "input_refinement_file" for refinement (raw data) and separately specify an input_map_file with a map and a data file with (optionally) detwinned data.) All the best, Tom T Hi, Detwinning is possible with xtriage or phenix.refine (on way or the other). Integrating this into density modification might be hard, especially when the twin fraction is high. What is your twin fraction? Tom, Is it possible to use a different set of amplitudes for density modification then for refinement? You wouldn't want to refine against detwinned data. Cheers Peter 2009/3/11 Thomas C. Terwilliger <terwilliger(a)lanl.gov>: > Hi Mary, > > I have not tried the phenix multi-crystal averaging procedure with > twinned > data, but here are some suggestions of how to proceed. > > 1. This routine does not do any refinement, only density modification. > The density modification step does not know about twinning, so it will > be > necessary (if your data are significantly twinned) to supply structure > factor files with detwinned data. I will check with Peter Zwart on > producing detwinned data in phenix; it may be necessary to do that with > other tools as normally phenix.refine uses the original data and does > the > calculations internally. > > 2. The models you supply are used to (a) identify NCS symmetry and > cross-crystal symmetry. Therefore it should not really matter if these > models are incomplete, and therefore the approach you are using should > work for improving the region where you have no model. The region where > NCS is applied is defined as a region centered near the center-of-mass > of > the supplied model over which the NCS correlation is high. > > > I hope that helps! > All the best, > Tom T > > > I started running this function and then realized that I probably need > to > do something to take the perfect twinning into account. I'm not sure of > the best way to do this. I believe the map coefficients output by a run > phenix.refine should be detwinned as I give a twin law when running > phenix.refine. Should I detwin the mtz containing the data and then use > those files for the multi crystal averaging? Is there a way to output > detwinned data mtz files in phenix or do I need to use another program? > > Also, I have two ncs copies in the ASU. There's a region of density in > both ncs copies that I'm trying to improve the maps for with the multi > crystal averaging. I don't have anything built in the region yet. Is > there a way to use the NCS and multi crystal averaging at the same time > without any pdb coordinates in that region? > > Even with my goof up about the twinning, the eff script I created > appeared to run without problems. > > Thanks, > > Mary X. Fitzgerald > Postdoctoral Associate > Arnold Lab > _______________________________________________ > phenixbb mailing list > phenixbb(a)phenix-online.org > http://www.phenix-online.org/mailman/listinfo/phenixbb > > > > > _______________________________________________ > phenixbb mailing list > phenixbb(a)phenix-online.org > http://www.phenix-online.org/mailman/listinfo/phenixbb > -- ----------------------------------------------------------------- P.H. Zwart Beamline Scientist Berkeley Center for Structural Biology Lawrence Berkeley National Laboratories 1 Cyclotron Road, Berkeley, CA-94703, USA Cell: 510 289 9246 BCSB: http://bcsb.als.lbl.gov PHENIX: http://www.phenix-online.org CCTBX: http://cctbx.sf.net -----------------------------------------------------------------

1 0

Re: [phenixbb] phenix.multi_crystal_average with twinned data
by Thomas C. Terwilliger 11 Mar '09

11 Mar '09

Hi Mary, I have not tried the phenix multi-crystal averaging procedure with twinned data, but here are some suggestions of how to proceed. 1. This routine does not do any refinement, only density modification. The density modification step does not know about twinning, so it will be necessary (if your data are significantly twinned) to supply structure factor files with detwinned data. I will check with Peter Zwart on producing detwinned data in phenix; it may be necessary to do that with other tools as normally phenix.refine uses the original data and does the calculations internally. 2. The models you supply are used to (a) identify NCS symmetry and cross-crystal symmetry. Therefore it should not really matter if these models are incomplete, and therefore the approach you are using should work for improving the region where you have no model. The region where NCS is applied is defined as a region centered near the center-of-mass of the supplied model over which the NCS correlation is high. I hope that helps! All the best, Tom T I started running this function and then realized that I probably need to do something to take the perfect twinning into account. I'm not sure of the best way to do this. I believe the map coefficients output by a run phenix.refine should be detwinned as I give a twin law when running phenix.refine. Should I detwin the mtz containing the data and then use those files for the multi crystal averaging? Is there a way to output detwinned data mtz files in phenix or do I need to use another program? Also, I have two ncs copies in the ASU. There's a region of density in both ncs copies that I'm trying to improve the maps for with the multi crystal averaging. I don't have anything built in the region yet. Is there a way to use the NCS and multi crystal averaging at the same time without any pdb coordinates in that region? Even with my goof up about the twinning, the eff script I created appeared to run without problems. Thanks, Mary X. Fitzgerald Postdoctoral Associate Arnold Lab _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org http://www.phenix-online.org/mailman/listinfo/phenixbb

1 0

phenix.multi_crystal_average with twinned data
by Mary Fitzgerald 11 Mar '09

11 Mar '09

I started running this function and then realized that I probably need to do something to take the perfect twinning into account. I'm not sure of the best way to do this. I believe the map coefficients output by a run phenix.refine should be detwinned as I give a twin law when running phenix.refine. Should I detwin the mtz containing the data and then use those files for the multi crystal averaging? Is there a way to output detwinned data mtz files in phenix or do I need to use another program? Also, I have two ncs copies in the ASU. There's a region of density in both ncs copies that I'm trying to improve the maps for with the multi crystal averaging. I don't have anything built in the region yet. Is there a way to use the NCS and multi crystal averaging at the same time without any pdb coordinates in that region? Even with my goof up about the twinning, the eff script I created appeared to run without problems. Thanks, Mary X. Fitzgerald Postdoctoral Associate Arnold Lab

1 0

March 15, 2009 deadline- User proposal submission for Collaborative Crystallography at BCSB
by Banumathi Sankaran 10 Mar '09

10 Mar '09

Dear Users, Deadline for May/June 2009 proposals will be March 15, 2009. Through the Collaborative Crystallography Pilot Program (CC) at the Advanced Light Source, scientists can send protein crystals to Berkeley Center for Structural Biology (BCSB) staff researchers for data collection and analysis. The CC Pilot 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. Thumbnail Description: ====================== The Collaborative Crystallography program will be piloted on beamlines 5.0.1 and 5.0.2 for one year; if successful it will be implemented on a permanent basis. CC proposals will go through the regular 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 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 submit a proposal, go to: http://www-als.lbl.gov/als/quickguide/independinvest.html Click on "protein crystallography" to begin proposal. For question 3 select "Collaborative PX 501/502." For question 9, please describe a specific research project with a clear end point. In order to request CC time for May/June 2009 allocation period, proposals must be submitted by March 15, 2009. The deadline for CC proposals for the time period July/August 2009 will be May 15, 2009. Regards, Banumathi Sankaran

1 0

ADP group definitions
by Roeland Boer 09 Mar '09

09 Mar '09

Dear all, I have a question regarding the definition of the ADP groups. I'm refining a structure using a different isotropic B for each domain, while applying TLS for some mobile domains (I'm also using NCS restraints for all equivalent domains). In the refinement log file, however, the isotropic B of the domains refined with TLS, as reported in the "TLS refinement" section, is equal to the B of the starting model in all refinement macro cycles. It seems, therefore, that the isoB for the TLS groups in not refined, although I've included these domains in the .adp.group definitions (the log file actually confirms that there are 7 adp groups). Is it possible to refine TLS and isotropic B's on the same part of a model? I include the input file below. Thanks for any suggestions, Roeland Boer. refinement.input.xray_data.labels=F refinement.refine.strategy=tls+individual_sites+group_adp refinement.refine.adp.tls="chain A and resid 4:132" refinement.refine.adp.tls="chain B and resid 4:132" refinement.refine.adp.tls="chain C and resid 4:132" refinement.refine.adp.tls="chain X and resid 4:132" refinement.refine.adp.tls="chain Y and resid 4:132" refinement.refine.adp.tls="chain Z and resid 4:132" refinement.refine.adp.one_adp_group_per_residue=False refinement.refine.adp.group="resid 133:203" refinement.refine.adp.group="chain A and resid 4:132" refinement.refine.adp.group="chain B and resid 4:132" refinement.refine.adp.group="chain C and resid 4:132" refinement.refine.adp.group="chain X and resid 4:132" refinement.refine.adp.group="chain Y and resid 4:132" refinement.refine.adp.group="chain Z and resid 4:132" refinement.main.number_of_macro_cycles=4 refinement.input.xray_data.low_resolution=20.0 refinement.input.xray_data.high_resolution=3.4 refinement.main.ncs = True refinement.ncs.find_automatically = False refinement.ncs.restraint_group { reference = chain A and resid 133:203 selection = chain B and resid 133:203 selection = chain C and resid 133:203 selection = chain X and resid 133:203 selection = chain Y and resid 133:203 selection = chain Z and resid 133:203 } refinement.ncs.restraint_group { reference = chain B and resid 4:132 selection = chain A and resid 4:132 selection = chain C and resid 4:132 selection = chain X and resid 4:132 selection = chain Y and resid 4:132 selection = chain Z and resid 4:132 } refinement.output.prefix=test

2 2

RuntimeError: Not enough memory for cubicles:
by Gino Cingolani 09 Mar '09

09 Mar '09

Dear Phenix community, I am still having problems to refine a large unit cell (~13,500 residues/asymmetric unit). The refinement goes well but at the very last cycle, immediately prior to outputting the refined model, phenix stops with the following error message: $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ ========== residual map mFobs-DFmodel: highest peaks and deepst holes ========= ----------peaks---------- Traceback (most recent call last): File "/usr/local/phenix-1.4-3/phenix/phenix/command_line/refine.py", line 9, in <module> command_line.run(command_name="phenix.refine", args=sys.argv[1:]) File "/usr/local/phenix-1.4-3/phenix/phenix/refinement/command_line.py", line 89, in run call_back_after_monitor_collect=call_back_after_monitor_collect) File "/usr/local/phenix-1.4-3/phenix/phenix/refinement/driver.py", line 1098, in run call_back_after_monitor_collect = call_back_after_monitor_collect) File "/usr/local/phenix-1.4-3/phenix/phenix/refinement/strategies.py", line 710, in refinement_machine fmodels=fmodels, model=model, log=log) File "/usr/local/phenix-1.4-3/phenix/phenix/refinement/strategies.py", line 738, in highest_peaks_and_deepest_holes_in_residual_map log = log) File "/usr/local/phenix-1.4-3/mmtbx/find_peaks.py", line 212, in show_highest_peaks_and_deepest_holes log = log) File "/usr/local/phenix-1.4-3/mmtbx/find_peaks.py", line 117, in __init__ map = fft_map_data).all(max_clusters = max_number_of_peaks) File "/usr/local/phenix-1.4-3/cctbx/maptbx/__init__.py", line 196, in peak_search max_clusters=parameters.max_clusters()) File "/usr/local/phenix-1.4-3/cctbx/maptbx/__init__.py", line 303, in __init__ general_positions_only=self._general_positions_only) File "/usr/local/phenix-1.4-3/cctbx/crystal/__init__.py", line 479, in site_cluster_analysis cubicle_epsilon=cubicle_epsilon) RuntimeError: Not enough memory for cubicles: This may be due to unreasonable parameters: cubicle_edge=0.7 space_span=(293.298,92.903,214.527) n_cubicles=(419,133,307) max_number_of_bytes=1724676096 Traceback (most recent call last): File "/usr/local/phenix-1.4-3/phenix/phenix/command_line/refine.py", line 9, in <module> command_line.run(command_name="phenix.refine", args=sys.argv[1:]) File "/usr/local/phenix-1.4-3/phenix/phenix/refinement/command_line.py", line 89, in run call_back_after_monitor_collect=call_back_after_monitor_collect) File "/usr/local/phenix-1.4-3/phenix/phenix/refinement/driver.py", line 1098, in run call_back_after_monitor_collect = call_back_after_monitor_collect) File "/usr/local/phenix-1.4-3/phenix/phenix/refinement/strategies.py", line 710, in refinement_machine fmodels=fmodels, model=model, log=log) File "/usr/local/phenix-1.4-3/phenix/phenix/refinement/strategies.py", line 738, in highest_peaks_and_deepest_holes_in_residual_map log = log) File "/usr/local/phenix-1.4-3/mmtbx/find_peaks.py", line 212, in show_highest_peaks_and_deepest_holes log = log) File "/usr/local/phenix-1.4-3/mmtbx/find_peaks.py", line 117, in __init__ map = fft_map_data).all(max_clusters = max_number_of_peaks) File "/usr/local/phenix-1.4-3/cctbx/maptbx/__init__.py", line 196, in peak_search max_clusters=parameters.max_clusters()) File "/usr/local/phenix-1.4-3/cctbx/maptbx/__init__.py", line 303, in __init__ general_positions_only=self._general_positions_only) File "/usr/local/phenix-1.4-3/cctbx/crystal/__init__.py", line 479, in site_cluster_analysis cubicle_epsilon=cubicle_epsilon) RuntimeError: Not enough memory for cubicles: This may be due to unreasonable parameters: cubicle_edge=0.7 space_span=(293.298,92.903,214.527) n_cubicles=(419,133,307) max_number_of_bytes=1724676096 $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ Following the various suggestions received after my previous email (~2 weeks ago), I have tried the following: --> Using phenix 1.4-3, which gave exactly the same problem as phenix 1.3 --> Using MACRO_CYCLE=0 before refinement to see if phenix can write out such a large asymmetric unit, and that works. Phenix does write out the model at a starting Rfree ~35.5%. --> Removing ordered_solvent=true Right now the refinement scheme I am using is as follows: phenix.refine XX.mtz model.pdb simulated_annealing=true elbow.XX_pdb.cif strategy=individual_sites+group_adp+tls tls_group_selections.params main.number_of_macro_cycles=5 main.ncs=true refinement.ncs.excessive_distance_limit=None refinement.pdb_interpretation.clash_guard.nonbonded_distance_threshold=No Once again, the refinement proceeds smoothly for 2-3 days; the Rfree drops nearly 3 points, and right at the very end, the programs stops without outputting a refined model. Does anybody have any idea what's going on? Thanks, Gino *********************************************************** Gino Cingolani, Ph.D. Assistant Professor Dept. of Biochemistry and Molecular Biology SUNY Upstate Medical University 750 E. Adams Street, Syracuse, NY, 13210 Tel. (315) 464 8744 Fax. (315) 464 8750 Email: cingolag(a)upstate.edu *********************************************************** "Nati non foste per viver come bruti, ma per seguir virtute e conoscenza" ("You were not born to live like brutes, but to follow virtue and knowledge") Dante, The Divine Comedy (Inferno, XXVI, vv. 119-120)

2 1

installation : cctbx or cci apps
by Bryan W. Lepore 09 Mar '09

09 Mar '09

i hope this is a "yes" or "no" question : i just got the latest phenix and the latest cci_apps running together. is the latest cctbx necessary? -bryan

2 1

Restraints on water molecules
by Engin Ozkan 08 Mar '09

08 Mar '09

Are restraints on water molecules overwritten on successive cycles of refinement where ordered solvent is on? I seem to lose my distance restraints between specific waters and metal, unless I name my water molecules something else (and supply phenix with a cif for that name which is identical to water inside). The distance restraints also seem to be in effect for all cycles if ordered_solvent is off. I am also trying to make sure water numbers don't change during solvent picking/deleting. Engin

2 3

PDF slides: introduction to refinement, phenix.refine and why automation is important (PHENIX)
by Pavel Afonine 08 Mar '09

08 Mar '09

Dear PHENIX community: I would like to draw your attention to these two PDF files with the slides that I recently made for a PHENIX workshop (workshop on Neutron crystallography, March 5-6, Japan). The first file (69 of very dense slides) makes a general introduction to crystallographic structure refinement (certainly biased towards how things are done in phenix.refine), plus some introduction to phenix.refine functionality. The second PDF file talks about PHENIX and "quick validation" with the strong accent of why automation is important. In general, I don't like and normally don't do such advertisements, but in this case I believe I can make an exception since I spent too much "of my software development" time on making these slides and they might be instructive for (new) PHENIX users. The links are (the first two!): http://www.phenix-online.org/presentations/neutron_japan_2009/ Thanks, Pavel. PS> Two slides in the first file are borrowed from Sasha Urzhumtsev, and 2-3 slides are borrowed from Paul Adams (second file).

2 2