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0 for wxc_scale and wxu_scale?
by Michaela Kramer, Biochemisches Inst. 26 Nov '08

26 Nov '08

I am refining a structure with a resolution of 2.1 Angstroem using Phenix. The spacegroup is P 31 2 1 (No. 152) with a twin fraction of 0.37 and twin law -h,-k,l. In the refinement procedure the NCS (4 chains) constraints are on, using individual sites, ADP and occupancy refinement. In the last refinement step, I use the option to optimize the values for wxu_scale and wxc_scale. These values are optimizes to 0 for wxc_scale and close to 0 for wxu_scale (0.2) When I tried this at earlier refinement stages wxu_scale was also here optimized in some refinement macrocycles to 0. Interestingly I obtain better R values (0.1948/0.2184), than without the optimization by using the default values (0.1391/0.2438 Rwork/Rfree). In addition to that the statistics are better: clashscore 14.41 (compared to 25.73 without wxc_scale & wxu_scale optimization), rotamer outliers 2.33% (compared o 4.66%) and Ramachandran favoured (Molprobity). How is it possible that wxu_scale and wxc_scale can be optimized to 0 by having better statistics? The total refinement target is defined as Etotal= wxc_scale x wxc x Exray + wc x Egeom As I understand it, the crystallographic refinement target (Exray) would now not be considered in the refinement procedure. Would this refinement procedure be valid? And how is wxc (output of target weights: x-ray data) defined? Thank you very much for your help Michaela Here is the output belonging to the optimization: Refinement Macrocycle 1 of 3 ========================== Target weights: x-ray data ========================= |-----------------------------------------------------------------------------| | XYZ refinement: T = Eexperimental * wxc * wxc_scale + Echem * wc | | wxc = 1.649809 wxc_scale = 0.500 wc = 1.000 | | angle between x-ray and geometry gradient vectors: 93.128 (deg) | | | | ADP refinement: T = Eexperimental * wxu * wxu_scale + Eadp * wu | | wxc = 0.888942 wxc_scale = 1.000 wc = 1.000 | | angle between Xray and ADP gradient vectors: 85.407 (deg) | |-----------------------------------------------------------------------------| ================================ xyz refinement =============================== Start r_free = 0.2118 scale= 0.0000 total_weight= 0.0000 r_free= 0.2181 0 scale= 1.0000 total_weight= 1.6498 r_free= 0.2336 0 scale= 1.5000 total_weight= 2.4747 r_free= 0.2379 1 scale= 2.0000 total_weight= 3.2996 r_free= 0.2412 2 scale= 0.6667 total_weight= 1.0999 r_free= 0.2297 0 scale= 0.5000 total_weight= 0.8249 r_free= 0.2245 1 scale= 0.4000 total_weight= 0.6599 r_free= 0.2226 2 Best r_free = 0.2181 ================================ ADP refinement =============================== ----------Individual ADP refinement---------- Start r_free = 0.2181 scale= 0.0000 total_weight= 0.0000 r_free= 0.2247 0 scale= 1.0000 total_weight= 0.8889 r_free= 0.2262 0 scale= 1.5000 total_weight= 1.3334 r_free= 0.2293 1 scale= 2.0000 total_weight= 1.7779 r_free= 0.2310 2 scale= 0.6667 total_weight= 0.5926 r_free= 0.2236 0 scale= 0.5000 total_weight= 0.4445 r_free= 0.2221 0 scale= 0.4000 total_weight= 0.3556 r_free= 0.2208 0 scale= 0.3333 total_weight= 0.2963 r_free= 0.2203 0 scale= 0.2857 total_weight= 0.2540 r_free= 0.2195 0 scale= 0.2500 total_weight= 0.2222 r_free= 0.2191 0 scale= 0.2222 total_weight= 0.1975 r_free= 0.2186 0 scale= 0.2000 total_weight= 0.1778 r_free= 0.2183 0 Best r_free = 0.2183 Refinement Macrocycle 2 of 3 ========================== Target weights: x-ray data ========================= |-----------------------------------------------------------------------------| | XYZ refinement: T = Eexperimental * wxc * wxc_scale + Echem * wc | | wxc = 1.737535 wxc_scale = 0.500 wc = 1.000 | | angle between x-ray and geometry gradient vectors: 89.612 (deg) | | | | ADP refinement: T = Eexperimental * wxu * wxu_scale + Eadp * wu | | wxc = 0.228155 wxc_scale = 1.000 wc = 1.000 | | angle between Xray and ADP gradient vectors: 155.392 (deg) | |-----------------------------------------------------------------------------| ================================ xyz refinement =============================== Start r_free = 0.2179 scale= 0.0000 total_weight= 0.0000 r_free= 0.2223 0 scale= 1.0000 total_weight= 1.7375 r_free= 0.2272 0 scale= 1.5000 total_weight= 2.6063 r_free= 0.2310 1 scale= 2.0000 total_weight= 3.4751 r_free= 0.2332 2 scale= 0.6667 total_weight= 1.1584 r_free= 0.2267 0 scale= 0.5000 total_weight= 0.8688 r_free= 0.2258 1 scale= 0.4000 total_weight= 0.6950 r_free= 0.2269 2 Best r_free = 0.2223 ================================ ADP refinement =============================== ----------Individual ADP refinement---------- Start r_free = 0.2223 scale= 0.0000 total_weight= 0.0000 r_free= 0.2228 0 scale= 1.0000 total_weight= 0.2282 r_free= 0.2235 0 scale= 1.5000 total_weight= 0.3422 r_free= 0.2256 1 scale= 2.0000 total_weight= 0.4563 r_free= 0.2275 2 scale= 0.6667 total_weight= 0.1521 r_free= 0.2219 0 scale= 0.5000 total_weight= 0.1141 r_free= 0.2213 0 scale= 0.4000 total_weight= 0.0913 r_free= 0.2207 0 scale= 0.3333 total_weight= 0.0761 r_free= 0.2204 0 scale= 0.2857 total_weight= 0.0652 r_free= 0.2201 0 scale= 0.2500 total_weight= 0.0570 r_free= 0.2197 0 scale= 0.2222 total_weight= 0.0507 r_free= 0.2195 0 scale= 0.2000 total_weight= 0.0456 r_free= 0.2194 0 Best r_free = 0.2194 Macrocycle 2 of 3 ========================== Target weights: x-ray data ========================= |-----------------------------------------------------------------------------| | XYZ refinement: T = Eexperimental * wxc * wxc_scale + Echem * wc | | wxc = 2.157676 wxc_scale = 0.500 wc = 1.000 | | angle between x-ray and geometry gradient vectors: 93.409 (deg) | | | | ADP refinement: T = Eexperimental * wxu * wxu_scale + Eadp * wu | | wxc = 0.228399 wxc_scale = 1.000 wc = 1.000 | | angle between Xray and ADP gradient vectors: 140.390 (deg) | |-----------------------------------------------------------------------------| ================================ xyz refinement =============================== Start r_free = 0.2163 scale= 0.0000 total_weight= 0.0000 r_free= 0.2189 0 scale= 1.0000 total_weight= 2.1577 r_free= 0.2299 0 scale= 1.5000 total_weight= 3.2365 r_free= 0.2340 1 scale= 2.0000 total_weight= 4.3154 r_free= 0.2341 2 scale= 0.6667 total_weight= 1.4385 r_free= 0.2282 0 scale= 0.5000 total_weight= 1.0788 r_free= 0.2255 1 scale= 0.4000 total_weight= 0.8631 r_free= 0.2249 2 Best r_free = 0.2189 ================================ ADP refinement =============================== ----------Individual ADP refinement---------- Start r_free = 0.2189 scale= 0.0000 total_weight= 0.0000 r_free= 0.2218 0 scale= 1.0000 total_weight= 0.2284 r_free= 0.2231 0 scale= 1.5000 total_weight= 0.3426 r_free= 0.2250 1 scale= 2.0000 total_weight= 0.4568 r_free= 0.2272 2 scale= 0.6667 total_weight= 0.1523 r_free= 0.2215 0 scale= 0.5000 total_weight= 0.1142 r_free= 0.2209 0 scale= 0.4000 total_weight= 0.0914 r_free= 0.2201 0 scale= 0.3333 total_weight= 0.0761 r_free= 0.2197 0 scale= 0.2857 total_weight= 0.0653 r_free= 0.2195 0 scale= 0.2500 total_weight= 0.0571 r_free= 0.2193 0 scale= 0.2222 total_weight= 0.0508 r_free= 0.2191 0 scale= 0.2000 total_weight= 0.0457 r_free= 0.2190 0 Best r_free = 0.2190 Michaela Kramer PhD student Institute of Biochemistry Winterthurerstrasse 190 8057 Zuerich Switzerland

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trouble in phenix.refine
by Huiying Li 26 Nov '08

26 Nov '08

I was running phenix.refine in tls mode with the following command line: phenix.refine mydata.mtz mymodel.pdb strategy=tls tls_group_selection.param mylibrary.cif against a 2.1A data set, P21211 SG, a homodimer in AU, totally 10 tls groups. The following errors occured and program stopped. I do not know how to fix the problem. 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 68, in run log=log) .... ... .... 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 Your help is highly appreciated. 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

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twin axis for p21
by HUA YUAN 25 Nov '08

25 Nov '08

Dear Colleagues, I'd like to share with you with a twinning puzzle that's been troubling me for a while. I have a ~2.2 A data merged in p212121 space group with unit cell: 41.8300 117.4490 134.4840 90.0000 90.0000 90.0000 Translational NCS was detected but structure solution was able to be obtained by phased molecular replacement as we have a Au derivative data set. Obtaining experimental phase or MR alone didn't work. However, with this model and data set, the refinement didn't work (with R-free ~40% or higher if with more cycles). Different twinning tests have been carried out and no sign of twinning. All possible p222 space groups have been tried but no luck or even worse. Considering that translational symmetry and perfect twinning may make things deceiving, the data was merged into p21 and a twin-refinement was performed in phenix.refine. After 3 cycles of rigid-body and group ADP, a R-free of ~30% was obtained and the output map looks OK. It looks like we got it. But then things get interesting when we tested other two axis as the p21 unique axis. With all the three choices plus corresponding twin-refinement, we could get a similar R-free (30%). Besides, the three maps all look OK except for one missing some side chains. Then the structure was solve in p1 and Xtriage was used with the hope that one axis would stand out. However, the RvsR for all three axis are almost the same. Did I miss anything here? Of course, we could pick up the one axis we like best and get the structure solved ^---^. However, this puzzle would eat me alive for a very long time. Best, Hua

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solvent flipping
by Mark Collins 24 Nov '08

24 Nov '08

Hi Is there a solvent flipping (like solomon, ccp4) rather than flattening option in phenix? Mark

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Windows version of Phenix
by Graeme Winter 21 Nov '08

21 Nov '08

Hi Folks, This is a potentially heretical question I know, but are there any plans to port Phenix to Windows in the future? Certainly having access to the command-line versions of the autosol and autobuild wizards would be excellent... Thanks, Graeme

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Question about overall_best history
by Mo Wong 20 Nov '08

20 Nov '08

Hi all, I'm sure this is in one of the log files (and written somewhere in the documentation), but I can't find which solve_*.mtz and which subsequent resolve_*.mtz were used to make overall_best_denmod_map_coeffs.mtz. Could someone please tell me where I may find it. Thank you

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Difference Fouriers using AutoSol
by Mo Wong 14 Nov '08

14 Nov '08

Hi, I'm a new Phenix user, so please forgive me if I ask an obvious question... I have pretty good SIRAS phases from one derivative, and calculating difference maps for 2 other derivatives show decent peaks above 5 sigma in both these maps. I've tried to follow the MIR example, but no sites are found in the two other derivatives (I thought Phenix calculates difference Fouriers for other derivatives as soon as it finds sites in one of the datasets - but it doesn't appear to in my case). Anyway, the following is my effeort at trying to use the SIRAS phases of the good derivative and the 5sigma peaks in the 2 other derivatives to phase a better map. Please could someone advise me if this script won't do what I'd like it to (i.e., calulate difference Fouriers for derviatives 2 and 3, refine the HA sites that I've included, and calculate phases): cell 100.000 100.000 100.000 90.000 90.000 90.000 # cell params resolution 5.5 # Resolution solvent_fraction 0.47 input_seq_file seq.dat ##### READ IN DERIV 1 SIRAS PHASES ###### input_phase_file solve_4.mtz input_phase_labels FP SIGFP PHIB FOM HLA HLB HLC HLD ############### DERIV 2 ################# input_file_list nat.sca hg.sca # Native and deriv 2 nat_der_list Native Hg # identify files by ha type inano_list noinano inano # say if ano diffs to be used f_prime -2.1 f_double_prime 9.5 n_ha_list 0 6 # number of heavy-atoms sites_file hg-sites.xyz run_list start # read in datafiles for dataset run_list read_another_dataset # about to start a new dataset here ############### DERIV 3 ################# input_file_list nat.sca xe.sca # Native and deriv 3 nat_der_list Native Xe # identify files by ha type inano_list noinano inano # say if ano diffs to be used f_prime -0.1 f_double_prime 3.5 n_ha_list 0 11 # number of heavy-atoms sites_file xe-sites.xyz ########################################## Many thanks!

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General user proposal deadline November 15
by Peter Zwart 13 Nov '08

13 Nov '08

Dear All, The deadline for general user proposals for PX beamtime at the ALS is comming up soon! At the BCSB, we have 5 beamlines for which you can apply for beamtime. At the Berkeley Center for Structural Biology (BCSB) we have recently completed a number of upgrades: Sector 8: Recent improvements include: * A high-accuracy microdiffractometer on BL 8.2.1, allowing for centering and viewing of very small crystals * Robotic automounters which can increase the screening throughput at the beamlines, and allows for the implementation of remote data collection. * A new large format (315mm active surface area) CCD detector on BL 8.2.1, facilitating data collection on large unit cell crystals Sector 5: Beamlines (5.0.1 and 5.0.3) now generate 1.5x10^11 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 8x10^11 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. Remote data collection for general users on sector 5 is currently being tested for a limited number of users. Please contact beamline staff for details. Please visit http://bcsb.lbl.gov/ for more details about the Center and its beamlines. If you'd like to apply for next round of beamtime (Jan-Feb) at the Advanced Light Source, please submit a General User proposal by November 15, 2008. 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 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 -----------------------------------------------------------------

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Autosol: cell keyword
by Page, Rebecca 13 Nov '08

13 Nov '08

Hi phenixers: Is there a way to use the Autosol Wizard with unmerged data from Scalepack? I can't get it to recognize the 'cell' keyword. Thanks in advance, Rebecca

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restricted simulated annealing
by Raja Dey 12 Nov '08

12 Nov '08

Dear Friends, How can I restrict simulated annealing in phenix on part of my molecule, say 2 chains out of 3 or part of a particular chain? Thanking you in advance for your reply... Raja

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