- phenixbb - phenix-online.org

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

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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 -----------------------------------------------------------------

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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 -----------------------------------------------------------------

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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

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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

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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

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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

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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)

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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

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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

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