- phenixbb - phenix-online.org

rosetta installation problems
by Mengbin Chen 04 Jun '13

04 Jun '13

Dear Phenix Users, I am interested in using Rosetta to do MR, but unfortunately I have encountered some problems in installing it: I was able to get to the step where "scons: done building targets." showed up, and set up the environment by typing setenv /Users/yanghai/Desktop/rosetta-3.5 into the shell. I also filled out the "path to rosetta" in "Preferences" under "PHENIX GUI", which I believe it is correct. However, when I was ready to run MR-rosetta, it kept saying "cannot locate Rosetta programs. You may set the path in PHENIX GUI under preferences->Automation, or define the environment variable PHENIX_ROSETTA_PATH (recommended)". Any help would be apprecited! Thank you in advance, Mengbin Chen -- Mengbin Chen Department of Chemistry University of Pennsylvania

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EG4 not in phenix dictionary?
by Yarrow Madrona 03 Jun '13

03 Jun '13

Hello, I have previously refined EG4 and EG2 (tetra and diethylene glycol) as ligands and these were in the standard phenix library. I was under the impression that phenix shares the library from the CCP4 monomer library which I know should have EG4. Can anyone help me with this. I guess the alternative is to generate a restraints file. -Yarrow -- Yarrow Madrona Graduate Student Molecular Biology and Biochemistry Dept. University of California, Irvine Natural Sciences I, Rm 2403 Irvine, CA 92697

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MAX IV Life Science Director position
by Derek Logan 03 Jun '13

03 Jun '13

Hi, If anyone fancies becoming Life Science Director at the world's brightest light source, look no further than here: http://www.lunduniversity.lu.se/o.o.i.s?id=24914&Dnr=547454&Type=E /Derek ________________________________________________________________________ Derek Logan tel: +46 46 222 1443 Associate Professor mob: +46 76 8585 707 Dept. of Biochemistry and Structural Biology www.cmps.lu.se<http://www.cmps.lu.se> Centre for Molecular Protein Science www.maxlab.lu.se/node/307<http://www.maxlab.lu.se/node/307> Lund University, Box 124, 221 00 Lund, Sweden

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Re: [phenixbb] Discrepancy between R-factors from phenix.refine vs phenix Generate "Table 1"
by Michael Thompson 01 Jun '13

01 Jun '13

As another follow up to this point, I have had a recent example where Xtriage reported that my intensity statistics indicated twinning (<|L|>=0.399 and <L^2>=0.227, alpha=0.4). After having refined the structure with the suggested twin law, and making composite omit maps along the way (using autobuild with no twin law), I ran model vs. data at the end of refinement and noticed that it reported R/Rfree that were about 10% higher than the R/Rfree from refinement, similar to what the original post described. I checked the model vs. data logfile and found that the L-test routine used by model vs. data was failing, so the subsequent analysis was carried out as if there was no twinning. So in my case, the crystal was twinned, but model vs. data was not able to perform the L-test so it was outputting incorrect R-factors. The error message at the top of the model vs. data log file is: Twin analysis failed: Python argument types in l_test.__init__(l_test, miller_index, double, space_group, bool, float, float, float, int) did not match C++ signature: __init__(_object*, scitbx::af::const_ref<cctbx::miller::index<int>, scitbx::af::trivial_accessor> miller_indices, scitbx::af::const_ref<double, scitbx::af::trivial_accessor> intensity, cctbx::sgtbx::space_group space_group, bool anomalous_flag, long parity_h, long parity_k, long parity_l, unsigned long max_delta_h) Twin analysis failed: Python argument types in l_test.__init__(l_test, miller_index, double, space_group, bool, float, float, float, int) did not match C++ signature: __init__(_object*, scitbx::af::const_ref<cctbx::miller::index<int>, scitbx::af::trivial_accessor> miller_indices, scitbx::af::const_ref<double, scitbx::af::trivial_accessor> intensity, cctbx::sgtbx::space_group space_group, bool anomalous_flag, long parity_h, long parity_k, long parity_l, unsigned long max_delta_h) Perhaps there are some other users who are seeing this behavior because the L-test fails within model vs. data. Mike ----- Original Message ----- From: "Nathaniel Echols" <nechols(a)lbl.gov> To: "PHENIX user mailing list" <phenixbb(a)phenix-online.org> Sent: Wednesday, May 29, 2013 5:18:57 PM GMT -08:00 US/Canada Pacific Subject: Re: [phenixbb] Discrepancy between R-factors from phenix.refine vs phenix Generate "Table 1" On Wed, May 29, 2013 at 4:13 PM, Sam Stampfer <Samuel.Stampfer(a)tufts.edu> wrote: > When I refined my model in phenix, I used the twin law h,-h-k,-l. I read in > the documentation that twinning can account for some of this discrepancy, > but that the program is supposed to take twinning into account if it will > lower the calculated R-work by more than 2%, which it doesn't seem to have > done (or there is some other problem with my data). Okay, the problem is that your data don't actually appear to be twinned. The automatic method used by phenix.model_vs_data (which is used internally for Table 1 and the validation GUI) only tries possible twin laws if the results of the "L test" show a suspicious distribution of intensities. Your data look fine, so it doesn't bother trying the twin laws. That the R-factors are lower when you refine with a twin law isn't necessarily indicative of the data actually being twinned - Garib Murshudov has looked into this in detail but I confess to being ignorant of the math (but I can probably dig up his paper on the subject if anyone is interested). However, I'm pretty sure the data are actually in a higher-symmetry space group. Will send details and new files off-list (probably tomorrow at this rate). I should probably change some of the programs and/or documentation to make it more clear what is being done internally, since it took me a bit of digging to realize what was going on. In general, though, always be very careful before running twinned refinement! I have seen several users do this by mistake when they really had higher symmetry. The maps will also be more model-biased when using twinned refinement, so it's good to avoid doing this unless absolutely necessary. -Nat _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org http://phenix-online.org/mailman/listinfo/phenixbb -- Michael C. Thompson Graduate Student Biochemistry & Molecular Biology Division Department of Chemistry & Biochemistry University of California, Los Angeles miket(a)chem.ucla.edu

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Re: [phenixbb] refinement of partly occupied water molecules?
by krimmer＠staff.uni-marburg.de 31 May '13

31 May '13

Dear Nathaniel, dear Pavel, thank you for your fast answers! > currently phenix.refine can add automatically only fully occupied > waters (I am not sure it will ever use atomic model for partially > occupied waters). If necessary, add partially occupied waters > yourself manually. Make sure they have properly set altlocs > (alternative location identifiers): non blank (say A and B). Is it current praxis to let phenix.refine freely refine the occupancy of water molecules (adding partially occupied water molecules in coot and don’t fix the occupancy in phenix)? If I do so, these water molecules are refined by phenix.refine with occupancies between 0.4-0.8. My resolution range was 1.1 – 1.4 Å. > The nightly builds have an option in phenix.refine to place ions > automatically based on a variety of criteria (still being tuned). > However, what you describe doesn't sound like any ion coordination > motif that I've read about. Could you please send us a picture that > shows the density in context? Some of the other readers with more > experience in "blobology" may recognize it. [if our list server > complains about the size of the message, just ignore it, I'll approve > it for posting] Attached you will find the image of the blob. I think it’s easily visible that the density blob is in the middle of the five-ring and that there is also a (smaller) blob in the middle of the six-ring. The water which occupies the blob was not yet refined with phenix, I just put it there to measure the distances to the carbon atoms. Best regards, Stefan

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gap between Wilson B and average B
by fn1＠rice.edu 30 May '13

30 May '13

Dear all, I have a structure at about 3.5 A, which has a wilson B of about 109 and has an average B factor from model is only 49 (I did not use TLS). I read from the previous posts that wilson B is sometimes smaller than average B, but in my case it is reversed. I look at the distribution of model B factors, they are in the range of 8 to 132. Is there any problem with the refinement of this structure? Thanks in advance for all advice! Regards, Fengyun

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refinement of partly occupied water molecules?
by krimmer＠staff.uni-marburg.de 30 May '13

30 May '13

Dear all, I have a question concerning the refinement of partly occupied water molecules: in some of my macromolecular crystal structures with resolutions between 1.1 - 1.4 Å, mainly in the second solvation shell round positive Fo-Fc electron density blobs are detectable which show after assignment of a water molecule to these blob and subsequent refinement with Phenix refine a good-looking calculated 2Fo-Fc electron density (round shape, the 2Fo-Fc map appears at a contour level of σ = 1.0 - 1.4), but there also occurs a small negative Fo-Fc electron density detectable inside the 2Fo-Fc density blob. The negative Fo-Fc electron density disappears if the occupancy of the water molecule is refined by Phenix or manually set lower than 100% occupancy. Therefore, I think these positions are partly occupied by water molecules, but I am not sure how I should handle it/how it is generally handled. I would be thankful for any advice and/or literature about this topic. My second question is concerning a density blob in perfect coordination distance to an indolyl group (perfectly located in the middle of the five-ring, with 3.4 Å distance to every of the five carbon atoms of the five-ring). What‘s the best way to find out which ion possibly occupies this position? I tried Na+ and Ca2+, but after refinement big negative Fo-Fc blobs appear in both cases (maybe the ion at this position is only partly occupied?). There is no anomalous signal detectable at this position (data collection wavelength was 0.91841 nm). After the refinement of a water molecule at this position, a positive Fo-Fc election density is still there. Thank you for your help! Stefan

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R-merge
by Mengbin Chen 29 May '13

29 May '13

Dear Phenix Users, I have R-merge values equal 0.000 from highest resolution shell (which corresponds to values larger than 1.000) in a data set processed by HKL2000, with redundancies higher than 10 (I know this may help in increasing R-merge). I am not sure if this is going to be a problem of Table 1 for the reviewers. Could you kindly drop me your suggestions? Thank you in advance, Mengbin Chen -- Mengbin Chen Department of Chemistry University of Pennsylvania

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phenix.refine using anomalous concepts for neutron data?
by Derek Logan 29 May '13

29 May '13

Hi, I'm doing a joint X-ray neutron refinement in Phenix and have run into some behaviour that puzzles me. I had not been paying any attention to the use or non-use of anomalous signal in the neutron dataset, as This is what I get if I let phenix.refine itself decide whether or not to use anomalous data: ================================= Neutron data ================================ F-obs: neutron.mtz:FO,SIGFO,DANO,SIGDANO,ISYM Miller array info: neutron.mtz:FO,SIGFO,DANO,SIGDANO,ISYM Observation type: xray.reconstructed_amplitude Type of data: double, size=16672 Type of sigmas: double, size=16672 Number of Miller indices: 16672 Anomalous flag: True Unit cell: (removed) Space group: P 21 21 21 (No. 19) Systematic absences: 0 Centric reflections: 1258 Resolution range: 29.22 1.89826 Completeness in resolution range: 0.785008 Completeness with d_max=infinity: 0.78486 Bijvoet pairs: 6832 Lone Bijvoet mates: 1750 Anomalous signal: 0.0876 Number of F-obs in resolution range: 16672 Number of F-obs<0 (these reflections will be rejected): 0 Number of F-obs=0 (these reflections will be used in refinement): 0 Refinement resolution range: d_max = 29.2200 d_min = 1.8983 and this is what I get if I forcibly switch off use of anomalous data in the phenix.refine GUI: ================================= Neutron data ================================ F-obs: neutron.mtz:FO,SIGFO,DANO,SIGDANO,ISYM Miller array info: neutron.mtz:FO,SIGFO,DANO,SIGDANO,ISYM Observation type: xray.reconstructed_amplitude Type of data: double, size=16672 Type of sigmas: double, size=16672 Number of Miller indices: 16672 Anomalous flag: True Unit cell: (removed) Space group: P 21 21 21 (No. 19) Systematic absences: 0 Centric reflections: 1258 Resolution range: 29.22 1.89826 Completeness in resolution range: 0.785008 Completeness with d_max=infinity: 0.78486 Bijvoet pairs: 6832 Lone Bijvoet mates: 1750 Anomalous signal: 0.0876 force_anomalous_flag_to_be_equal_to=False Reducing data to non-anomalous array. R-linear = sum(abs(data - mean(data))) / sum(abs(data)) R-square = sum((data - mean(data))**2) / sum(data**2) In these sums single measurements are excluded. Redundancy Mean Mean Min Max Mean R-linear R-square unused: - 29.2234 bin 1: 29.2234 - 4.0863 1 2 1.699 0.0240 0.0011 bin 2: 4.0863 - 3.2448 1 2 1.793 0.0302 0.0022 bin 3: 3.2448 - 2.8350 1 2 1.800 0.0417 0.0041 bin 4: 2.8350 - 2.5760 1 2 1.764 0.0486 0.0054 bin 5: 2.5760 - 2.3914 1 2 1.745 0.0505 0.0051 bin 6: 2.3914 - 2.2505 1 2 1.717 0.0513 0.0051 bin 7: 2.2505 - 2.1378 1 2 1.658 0.0533 0.0056 bin 8: 2.1378 - 2.0448 1 2 1.614 0.0611 0.0074 bin 9: 2.0448 - 1.9661 1 2 1.575 0.0634 0.0075 bin 10: 1.9661 - 1.8983 1 2 1.485 0.0736 0.0097 unused: 1.8983 - Fobs statistics after all cutoffs applied: Miller array info: None Observation type: xray.amplitude Type of data: double, size=9840 Type of sigmas: double, size=9840 Number of Miller indices: 9840 Anomalous flag: False Unit cell: (removed) Space group: P 21 21 21 (No. 19) Systematic absences: 0 Centric reflections: 1258 Resolution range: 29.22 1.89826 Completeness in resolution range: 0.855801 Completeness with d_max=infinity: 0.855503 What's going on here? If I let phenix.refine decide automatically, it looks like it's reading in F+ and F- separately and regarding them as independent in the refinement. If I force "no anomalous" it merges them. However I can't work out whether in either case it has actually read FOBS instead of the Friedel mates. According to the SCALA manual the I+ and I- columns will always be written out even if the keyword ANOMALOUS OFF is used, so this is a potential pitfall for many. In addition, phenix.refine seems to identify the neutron data first as xray.reconstructed_amplitude then as xray.amplitude even though the data are explicitly defined as neutron data in the GUI. I can send more information if required! Thanks Derek ________________________________________________________________________ Derek Logan tel: +46 46 222 1443 Associate Professor mob: +46 76 8585 707 Dept. of Biochemistry and Structural Biology www.cmps.lu.se<http://www.cmps.lu.se> Centre for Molecular Protein Science www.maxlab.lu.se/node/307<http://www.maxlab.lu.se/node/307> Lund University, Box 124, 221 00 Lund, Sweden www.saromics.com<http://www.saromics.com>

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Structure Comparison questions
by Heather Condurso 29 May '13

29 May '13

Dear all, I have a few related structures that I would like to compare to make figures for publication but I have a few issues. The biggest issue is that my structures are large (12- 422aa chains) and any attempt at simple alignment in pymol results in a crash. I would like to first compare entire structures, but also compare the differences between related chains in the same structure. I want to look at the small differences in the active site density. What is the best way to do this? Is there a simple way to do this or do I need to make separate pdb files for each chain? Can I use the same mtz or should I rename copies? I will try a few things but any advice would be greatly appreciated. Heather condurso(a)bc.edu

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R: problem refining RNA structure at 4Å resolution
by fulvio.saccoccia＠uniroma1.it 29 May '13

29 May '13

Any signs of twinning? Check also the correctness of your space group: probably you could index In a lower symmetry Laue group with a slight increased distorsion index. Fulvio ----Messaggio Originale---- Da: Benoît Masquida Inviato: 29/05/2013, 14:53 A: PHENIX user mailing list Oggetto: [phenixbb] problem refining RNA structure at 4Å resolution Dear phenix users, I solved using MR a low resolution RNA structure taking data up to 4.3Å and a model of a structure with slight differences. However, when I try to refine, Rwork decreases quickly (start 47, final 32 but Rfree remains high (start 48, final 44). The data are pretty good up to 4.0 with I/sigma above 1. The map doesn't point clearly to wrong regions of the model. What can I try to improve the maps and build the correct model? Thanks in advance. Benoit _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org http://phenix-online.org/mailman/listinfo/phenixbb

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problem refining RNA structure at 4Å resolution
by Benoît Masquida 29 May '13

29 May '13

Dear phenix users, I solved using MR a low resolution RNA structure taking data up to 4.3Å and a model of a structure with slight differences. However, when I try to refine, Rwork decreases quickly (start 47, final 32 but Rfree remains high (start 48, final 44). The data are pretty good up to 4.0 with I/sigma above 1. The map doesn't point clearly to wrong regions of the model. What can I try to improve the maps and build the correct model? Thanks in advance. Benoit

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Re: [phenixbb] paired refinement in phenix gui
by Kay Diederichs 28 May '13

28 May '13

On Mon, May 27, 2013 at 15:08:26 -0700, Nat Echols wrote: >> >It would be nice if you could confirm if my settings are ok, or if there are >> >still some settings in the background which I overlooked, because we see >> >small changes in R and Rfree. > Changes relative to what? It is not expected that feeding the output > PDB file back to phenix.refine (or another program) will exactly > reproduce the R-factors from the previous round of refinement, because > the precision of the atomic parameters is limited by PDB format. > (Using mmCIF instead would be more reliable, but we don't think a > change in R-factor of 0.0004 is worth worrying about anyway.) > >> >I added the model refined at higher resolution (e.g. 1.5A) and the mtz file >> >cut at CC (e.g.1.5A) in the ?Input data? tab, and set High resolution cutoff >> >in ?X-ray data and experimental phases? box to (e.g. 1.8A) >> >In the refinement setting strategy tab I uncheck everything and set Number >> >of cycles = 1. >> >In ?other options? I uncheck everything too. > This sounds correct, but it's much easier to simply run the validation > GUI, because the underlying code (specifically phenix.model_vs_data) > is designed to calculate identical R-factors to what phenix.refine > shows you. From what I've seen, recent phenix.model_vs_data works correctly and reproduces the statistics from phenix.refine within the numerical accuracy that one can expect. > >> >Alternatively adding a file to the input data with > I don't recommend this - using parameter files as input is fine for > large repetitive parameter blocks such as custom geometry restraints, > selections from the TLSMD server, or nucleic acid base pairs, but > anything that attempts to override the controls in the main GUI may > not work. > >> >2. For whatever reason I would like to do that (molprobity) - can I (and how >> >can I do this) add the command line commands also in the gui? > Could you please clarify? > >> >3. In december 2012 there was a discussion about weak data and refinement. I >> >would like to know if there are new data available showing that ?Whether the >> >benefit they describe is considered cosmetic or non-trivial,.....? > Maybe Kay could comment on this... There is not a single answer that fits all situations. When done correctly (e.g. keeping Rfree flags, keeping refinement strategy), in those cases that I looked at, using weak reflections (cutoff at <I/sigma>~0.5 to 1, cc1/2~0.15-0.25) improved the model's Rfree at lower resolution (paired refinement!) by a few percent, compared to cutting the data at I/sigma>2. Just try it and see if the improvement is worth it, according to your own judgement! > >> >4. Additionally could one short comment on what phenix is doing with strong >> >(weak) reflections marked as aliens in XDS. > Assuming that XDS doesn't actually remove these, phenix.refine may or > may not discard them at the outlier filtering stage (which can be > disabled). I don't know how the different methods for flagging > suspect resolutions compare; Kay and/or Pavel would know more about > this. The "Alien" list in CORRECT.LP was only (originally) meant to alert the user to ice reflections, zingers, and cosmic rays (not sure if the latter play a role, but the former certainly do!). If the "Alien" list just shows reflections at very high resolution, in low-<intensity> resolution shells, then this is due to the fact that Wilson statistics does not hold in these shells, and therefore the algorithm for deciding about "Aliens" is no longer valid. If this happens, you should (as identified in analysis together with James Holton) prevent CORRECT from marking "Aliens" as outliers by changing REJECT_ALIEN from 20 (the default) to 100 or so. Otherwise the mean intensity in that resolution shell will drop (due to rejecting the strongest reflections), and that is bad for applying the Wilson&French algorithm. The XDSCONV in the latest XDS package (identifying itself as March-30-2013) has two fixes for dealing with weak data (again, based on an exchange with James Holton). Unfortunately, a small bug (affecting very few datasets) was identified by Graeme Winter (thanks!) in the XDS program of that package, so be prepared to download the XDS package again in a few days. hope that helps, Kay > >> >According to the supplementary of P. Andrew Karplus, Kay Diederichs ?Linking >> >Crystallographic Model and Data Quality? >> >I would like to do >> >anisotropic refinement, with additional options >> >?ordered_solvent.new_solvent=anisotropic >> >adp.individual.anisotropic="not element H" >> >fix_rotamers=True?. > If your resolution really is 1.5?, I wouldn't recommend making waters > anisotropic. There is some difference of opinion whether anisotropic > refinement of protein atoms is acceptable; it's probably good to try > both ways. (It would be interesting to know whether including the > weak high-resolution data allows you to parameterize the model more > aggressively.) > > -Nat > -- Kay Diederichs http://strucbio.biologie.uni-konstanz.de email: Kay.Diederichs(a)uni-konstanz.de Tel +49 7531 88 4049 Fax 3183 Fachbereich Biologie, Universität Konstanz, Box M647, D-78457 Konstanz This e-mail is digitally signed. If your e-mail client does not have the necessary capabilities, just ignore the attached signature "smime.p7s".

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PDB deposition
by Kay Diederichs 27 May '13

27 May '13

Dear Phenix developers, the following questions came up today, during an attempt to answer the questions that the PDB asks at deposition time: a) For Bulk solvent modeling, what should be given as value for K sol and B sol ? My understanding is that the new solvent model does no longer result in these values; the table is Total model structure factor: F_model = k_total * (F_calc + k_mask * F_mask) k_total = k_isotropic * k_anisotropic Resolution Compl Nwork Nfree R_work <Fobs> <Fmodel> kiso kani kmask 37.014-13.525 85.34 95 4 7.2146 38.755 318.867 0.937 1.000 0.300 13.240-10.663 91.82 91 10 7.1453 40.328 328.027 0.901 1.000 0.309 10.640-8.570 93.27 202 6 7.4011 38.069 318.901 0.905 1.002 0.310 8.548-6.875 95.70 380 21 7.2679 31.166 258.647 0.901 1.000 0.309 6.871-5.523 95.40 731 36 7.1964 29.485 241.669 0.908 1.001 0.302 5.520-4.439 90.49 1347 61 7.3445 39.504 329.514 0.934 1.001 0.281 4.438-3.568 92.52 2610 150 7.3166 41.793 347.063 0.961 1.000 0.280 3.567-2.867 94.10 5191 231 7.2983 33.593 278.704 0.965 1.000 0.024 2.867-2.304 92.97 9835 476 7.3098 23.922 198.755 0.984 1.000 0.000 2.304-1.852 92.81 18869 947 7.3097 18.746 155.751 1.053 1.000 0.000 1.852-1.488 90.33 35341 1797 7.3288 10.914 90.931 1.035 0.999 0.000 1.488-1.196 86.49 64979 3561 7.2975 6.848 56.810 1.028 0.998 0.000 1.196-1.081 53.28 29672 1567 7.1776 5.351 43.737 0.986 1.000 0.000 Approximation of k_total with k_overall*exp(-b_overall*s**2/4) k_overall=1.0149 b_overall=-0.0072 So what can we do here? b) there is the question about Overall Anisotropic values B11; B22; B33; B12; B13; B23.... but the logfile offers overall anisotropic scale matrix: V0: -0.0214,0.0415,-0.0249,0.0006,-0.0515,-0.0161 V1: -0.0055,-0.0022,0.0080,-0.0001,-0.0020,0.0011 So which of these should be given? I've tried to locate the information about these two questions in the documentation, but it seems I missed it! thanks, Kay

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command line molprobity tools
by Engin Özkan 27 May '13

27 May '13

Hi everybody, I want to create a comprehensive, human-readable log file by running validation using molprobity tools within phenix command-line. I do not have structure factors/maps. Apparently, the GUI runs validation through phenix.model_vs_data (according to the .eff file), but in command-line, I cannot get phenix.model_vs_data to run without an mtz/hkl. I guess I could provide it a fake data file, and then set the comprehensive flag to "True"? BTW, the log file produced by the GUI is not comprehensive at all. I could not find a related mmtbx or cctbx function either. Another mentioned tool, phenix.validate, does not even exist in my installation (dev 1361). Thanks, Engin -- Engin Özkan Post-doctoral Scholar Laboratory of K. Christopher Garcia Howard Hughes Medical Institute Dept of Molecular and Cellular Physiology 279 Campus Drive, Beckman Center B173 Stanford School of Medicine Stanford, CA 94305 w ph: (650)-498-7111 cell: (650)-862-8563

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Fwd: [ccp4bb] "Formageddon" is upon us... Important news from wwPDB!
by Paul Adams 24 May '13

24 May '13

Begin forwarded message: > From: Gerard DVD Kleywegt <gerard(a)XRAY.BMC.UU.SE> > Subject: [ccp4bb] "Formageddon" is upon us... Important news from wwPDB! > Date: May 24, 2013 11:20:51 AM PDT > To: CCP4BB(a)JISCMAIL.AC.UK > Reply-To: info(a)wwpdb.org > > Dear colleagues, > > I would like to draw your attention to a notification from the wwPDB partners about "Deposition and Release of PDB Entries Containing Large Structures" - see: > > http://www.wwpdb.org/news/news_2013.html#22-May-2013 > > There are major changes afoot in the way large structures are handled in the PDB, as well as in the deposition and annotation procedures and software used by the wwPDB sites. This is of immediate relevance for depositors and users of large structures, but also for software developers and anyone who routinely processes the entire PDB archive or its weekly releases (bioinformatics resources, etc.). From 2014, it will affect essentially everyone who deposits, uses or processes PDB entries. > > If you have any questions about the new deposition system or the procedures for handling large structures or any of the other changes, please contact: info(a)wwpdb.org > > Please pass on this information to anyone likely to be affected by the upcoming changes. Thanks! > > > On behalf of the Worldwide Protein Data Bank, > > --Gerard Kleywegt > > --- > Gerard J. Kleywegt, PDBe, EMBL-EBI, Hinxton, UK > gerard(a)ebi.ac.uk ..................... pdbe.org > Secretary: Pauline Haslam pdbe_admin(a)ebi.ac.uk -- Paul Adams Deputy Division Director, Physical Biosciences Division, Lawrence Berkeley Lab Division Deputy for Biosciences, Advanced Light Source, Lawrence Berkeley Lab Adjunct Professor, Department of Bioengineering, U.C. Berkeley Vice President for Technology, the Joint BioEnergy Institute Laboratory Research Manager, ENIGMA Science Focus Area Building 64, Room 248 Building 80, Room 247 Building 978, Room 4126 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: Louise Benvenue [ LBenvenue(a)lbl.gov ][ 1-510-495-2506 ] --

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Invitation to Participate in a NSF-funded workshop/conference
by Sibo Jiang 24 May '13

24 May '13

Invitation to Participate in a NSF-funded workshop/conference Dear Colleagues in Structural Biology, Recently we have received a NSF grant helping to build the community networks and find solutions for the challenging problems. We have also discussed with the program officer of US National Science Foundation on planning for a research conference/workshop series proposal to US NSF. This conference/workshop series will particularly focus on the technical issues and practical challenges in structural biology of nucleic acids, protein-nucleic acid complexes and small molecule-nucleic acid complexes. These challenges include crystallization, derivatization, phasing and high-resolution structure determination. You are invited to participate in this conference/workshop series. If you are interested in our invitation, please reply this email or send your email to huang(a)gsu.edu<mailto:[email protected]>. In your email, please let us know your name, research interests (up to five key words), affiliation and phone number. The details will be announced later. Colleagues in academia are encouraged to forward this email to your friends and colleagues in industry. Colleagues in industry are particularly welcome. Sincerely, The Planning Committee of the NSF-funded Conference/Workshop

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FW: need suggestion
by afshan490＠yahoo.com 24 May '13

24 May '13

<br/><br/><br/>Sent from Yahoo! Mail for iPhone

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From Dmitri Svergun and Matthias Wilmanns: 11th International Conference: Biology and Synchrotron Radiation (BSR)
by Terwilliger, Thomas C 24 May '13

24 May '13

11th International Conference: Biology and Synchrotron Radiation (BSR) 8-11 September 2013, EMBL Hamburg, Germany Abstract submission deadline is approaching – 1st June 2013! Join us in Hamburg, Germany for BSR 2013! Scientists at all possible career levels are invited to this meeting. Check out our preliminary programme to learn more! http://www.bsr2013.org/BSR13-01/programme/index.html Young researchers are highly welcome to apply for a travel grant from the International Union of Crystallography when registering. http://www.bsr2013.org/BSR13-01/info/index.html Present your work! Selected talks and poster sessions provide great opportunities for participants to present their work. Unique networking opportunities are also available to meet colleagues, friends and future collaboration partners from all over the world. Discover the city of Hamburg! BSR 2013 includes a social programme with boat trips and day and night excursions. Accompanying persons will enjoy special rates! http://www.bsr2013.org/BSR13-01/attractions/index.html A tour of DESY’s most impressive research infrastructures - including PETRA, FLASH and the future X-FEL – is included in the registration fee. http://www.bsr2013.org/BSR13-01/desy/index.html Dates at a glance: · Registration and abstract submission: 1 June 2013 · Registration without abstract: 1 August 2013 Take a look at the updated BSR webpage (www.bsr2013.org<http://www.bsr2013.org>) for more information and to register. CONTACT Diah.Yulianti(a)embl.de<mailto:[email protected]> Alternatively, contact us on the event facebook page https://www.facebook.com/events/332259356868854/

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Call for Proposals: IMAGINE, a New Neutron Crystallography Diffractometer
by Meilleur, Flora 23 May '13

23 May '13

IMAGINE, Neutron Crystallography Diffractometer High Flux Isotope Reactor (HFIR), Oak Ridge National Laboratory Call for proposals for experiments anticipated to run from August through December 2013 You are invited to apply for beam time on the neutron quasi-Laue diffractometer IMAGINE, at the High Flux Isotope Reactor. Proposal will be accepted via the web-based proposal system until NOON Wednesday, July 31, 2013. This call is for experiments anticipated to run from August through December 2013. Please see the attached flyer for additional information. For technical information about the capabilities of IMAGINE go to neutrons.ornl.gov/imagine/<http://neutrons.ornl.gov/imagine/> or contact Flora Meilleur, meilleurf(a)ornl.gov, or Andrey Kovalevsky, kovalevskyay(a)ornl.gov. Flora Meilleur, Ph. D Assistant Professor, Molecular and Structural Biochemistry North Carolina State University IMAGINE lead scientist, Neutron Sciences Directorate Oak Ridge National Laboratory Phone: 865-242-5747

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conflicting bond_simple restraints
by Jianxun Qi 23 May '13

23 May '13

Hi, I had a phosphoserine (SEP) in my structure that is attached in this message. The data_link file for SEP already exists in the mon_lib (data_link_SEPC-N) and the corresponding cif_link I used is as follow: refinement.pdb_interpretation { apply_cif_link { data_link = "SEPC-N" residue_selection_1 = "chain C and resname PHE and resid 3" residue_selection_2 = "chain C and resname SEP and resid 4" } apply_cif_link { data_link = "SEPC-N" residue_selection_1 = "chain C and resname SEP and resid 4" residue_selection_2 = "chain C and resname ARG and resid 5" } } When I run the phenix.refine (my version is phenix-1.8-1069), it said conflicting bond_simple restraints: Sorry: Conflicting bond_simple restraints: 1. definition from: residues: PHE + SEP 2. definition from: residues: PHE + SEP atoms: "ATOM 3121 C PHE C 3 .*. C " "ATOM 3192 N SEP C 4 .*. N " Has anyone ever experienced this case before? Any hints are welcome. Regards Jianxun Qi May 22,2013

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Ca occupancy in multiple soaked structures
by Daniel Slade 22 May '13

22 May '13

Hi Everyone, I have a complex case of data analysis I would love some opinions on. I have a group of structures of a protein that binds 6 calciums, and we want to know the order/concentration at which they fill. The crystals were soaked with calcium from 0-10 mM and all are 1.6-2.1 A in resolution. I determine the order of the calcium binding, and now I am working on the percent occupancy at each concentration. To do this, I have removed the calciums, and then performed simulated annealing for each structure from 2.1-30A resolution to 'normalize' them. I then look at the omit map and scroll out to find the peak sigma intensity of where the calciums should be. The curves fit very well for each calcium, and the numbers correspond well with other solution studies we have. My thought is that I need to normalize this in another way, so I thought choosing a water molecule that is present in all of the structures and removing it, then performing simulated annealing would be a good idea to normalize the peak intensity for each calcium soak concentration, since this water molecule shouldn't change occupancy. Is this the proper way to do this, or can you guys suggest a better way to go about this. I also looked at B factor and e/A3, but that isn't as consistent as peak sigma. Thanks in advance. Dan Daniel J. Slade, Ph.D. Research Associate Thompson Lab The Scripps Research Institute 130 Scripps Way Jupiter, FL 33458 (561) 228-3213

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43rd Mid-Atlantic Macromolecular Crystallography Meeting -- reminder
by Charles Pemble 22 May '13

22 May '13

*43**rd** Mid-Atlantic Macromolecular Crystallography Meeting* *May 30 – June 1, 2013* *Duke University* *Durham, NC* This is a reminder for the 43rd Mid-Atlantic Macromolecular Crystallography Meeting, which will be held next week, May 30-June 01 at Duke University in Durham, North Carolina. Registration and details can be found at http://www.mid-atlantic.org. We encourage you to submit your abstracts for a poster presentation as the deadline is fast approaching! *DEADLINES (updated):* Abstract submission for poster - Next Monday, May 27th *No deadline for registration With best wishes on behalf of the primary Organizing Committee, Charlie Pemble _____________________________ Charles W. Pemble IV, Ph.D. Duke University Medical Center 308 Research Drive LSRC, A06 Durham, NC 27708 charles.pemble(a)duke.edu We thank our current sponsors for their generous contributions: Rigaku Americas Corporation, Agilent Technologies Inc., GlaxoSmithKline, Art Robbins, Bruker, HKL Research Inc., Molecular Dimensions, Hampton Research, Labcyte, GenScript, Genentech, Emerald BioSystems, TTP LabTech, Oxford Cryosystems, PhD Posters, Duke University School of Medicine, Duke Human Vaccine Institute, and Avanti Polar Lipids Inc.

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Workshop - The Future of Microfocus Protein Crystallography - 24-25 September, 2013, Lund, Sweden
by Marjolein Thunnissen 22 May '13

22 May '13

Dear all, I would like to announce a workshop on "The Future of Microfocus Protein Crystallography" which will take place at the MAX IV User meeting on the 24th and the 25th of September, 2013, in Lund, Sweden. Recent developments within protein crystallography methods in combination with the possibilities offered by the new upcoming synchrotron sources such as the MAX IV 3 GeV storage ring make possible exciting new ways of carrying out research within structural biology. Small microfocus beamlines with a very high flux and increased coherence will make new types of experimentation possible. The scientific possibilities will be highlighted through a series of talks by experts showing the impact of the new type of X-ray sources and microfocus beamlines on research within structural biology. The new generation of microfocus beamlines will also set a high demand on instrumentation and suitable software. For example, data collection will potentially be very fast (in milliseconds) and radiation damage will allow only a limited amount of data to be collected from each sample. New technical solutions for sample handling and presentation, data processing methods and more will be needed for these types of experiments, and these will also be presented and discussed at the workshop. Confirmed speakers include David Stuart, Daniel Wacker, Simone Weynand, Jens Preben Morth, Tove Sjögren, Richard Neutze, Janet Smith, Alke Meentz, Alexei Soares, Thomas White and Clemens Schulze-Briese The workshop is part of a process to obtain an updated proposal for the possible Phase 2b beam line MicroMAX at the upcoming new 3 GeV storage ring of MAX IV in Lund. This workshop should give the user community an opportunity to explore the scientific possibilities of a new generation of microfocus beam lines, but also it will give them the chance to contribute with specific input on their own experimental demands on the beam line. The workshop is part of the MAX IV laboratory user meeting. Registration will be through the user meeting website which will open for registration soon. Further information on this workshop can be found at https://www.maxlab.lu.se/node/1609 and for the MAX IV user meeting at https://www.maxlab.lu.se/um13 . For the organisers, Richard Neutze, Gunter Schneider and Gregers Rom Andersen Marjolein Thunnissen ________________________________________________________________________ Marjolein Thunnissen Phone +46-(0)76-632 0417 Associate Professor Fax +46-(0)46-22 24692 Dept of Biochemistry and Structural Biology, Lund University http://www.mps.lu.se<http://www.mps.lu.se/> PO-Box 124 S-221 00 Lund, Sweden Scientific coordinator MX (The MAX IV Laboratory): I911 and BioMAX ________________________________________________________________________

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depositing pdb files
by Mengbin Chen 21 May '13

21 May '13

Hi Phenix Users, I am going to deposit some pdb files into RCSB, which requires Luzzati plots and a bunch of other things. I don't know how to get these results from Phenix (actually the main problem is luzzati errors, Phenix does not seem to provide them), although SFcheck from CCP4i provides only some of them. I know CNS can do this by model_stats.inp, but that's too much trouble since I have complicated ligands bound. Any suggestions would be appreciated! Thank you in advance, Mengbin -- Mengbin Chen Department of Chemistry University of Pennsylvania

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mr_rosetta: how to continue from recovered mr_protocols.default run
by Christian Becke 21 May '13

21 May '13

Hi, I am trying mr_rosetta on a complex. At the GROUP_OF_ROSETTA_REBUILD stage, my machine ran out of memory and the mr_protocols.default processes were killed. After adding more swap space, I restarted the mr_protocols.default runs (3 parallel jobs, one for each chain) using the scripts RUN_?/WORK_?/run_rebuild.sh, which appear to have finished correctly (i.e., there are 20 pdb files generated for each of the chains). However, I could not figure out how to restart mr_rosetta so that it picks up these models. Any suggestions? Thanks, Christian

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Grid size in max_entropy maps
by Schubert, Carsten [JRDUS] 20 May '13

20 May '13

Is there a way to control the grid size in the maximum entropy maps? The current grid size is quite small, resulting in a very fine mesh. Thanks Carsten

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43rd Mid-Atlantic Macromolecular Crystallography Meeting
by Charles Pemble 10 May '13

10 May '13

*43**rd** Mid-Atlantic Macromolecular Crystallography Meeting* *May 30 – June 1, 2013* *Duke University* *Durham, NC* *ANNOUNCEMENT: *We have extended the deadline for submitting abstracts to be considered for presentations until midnight Sunday, May 12. Registration and details can be found at http://www.mid-atlantic.org. The 43rd Mid-Atlantic Macromolecular Crystallography Meeting, will be held at Duke University in Durham, North Carolina. This year will feature keynote speaker Tom Terwilliger from Los Alamos National Lab, workshops on PHENIX and MolProbity, plus many talks & posters on new and exciting research. The best presentation and poster by a Graduate Student & Postdoctoral Fellow will be awarded (total of 4). We encourage you to submit your abstracts and sign up for the workshops as these are separate processes from registration. *DEADLINES (updated):* Abstract submission for presentation - May 12th Abstract submission for poster - May 27th *No deadline for registration With best wishes on behalf of the primary Organizing Committee, Charlie Pemble _____________________________ Charles W. Pemble IV, Ph.D. Duke University Medical Center 308 Research Drive LSRC, A06 Durham, NC 27708 charles.pemble(a)duke.edu We thank our current sponsors for their generous contributions: Rigaku Americas Corporation, Agilent Technologies Inc., GlaxoSmithKline, Art Robbins, Bruker, HKL Research Inc., Molecular Dimensions, Hampton Research, Labcyte, GenScript, Genentech, Emerald BioSystems, TTP LabTech, Oxford Cryosystems, PhD Posters, Duke University School of Medicine, Duke Human Vaccine Institute, and Avanti Polar Lipids Inc.

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Submissions for next CCN newsletter
by Nigel Moriarty 07 May '13

07 May '13

Folks Calling for articles and short communications of interest to structural biologists. The deadline for submission in the June 2013 issue is 31 May, 2013 but if you can send me drafts before the 15th of May we can get started on generating proofs. The Computational Crystallography Newsletter (CCN) is an electronic newsletter for structural biologists, and is published online every 6 months. Feature articles, meeting announcements and reports can be submitted to me at any time for consideration. Submission of text by email or word-processing files using the CCN templates is requested. Past newsletters and a Microsoft Word template are available at www.phenix-online.org/newsletter. Cheers Nigel -- Nigel W. Moriarty Building 64R0246B, Physical Biosciences Division Lawrence Berkeley National Laboratory Berkeley, CA 94720-8235 Phone : 510-486-5709 Email : NWMoriarty(a)LBL.gov Fax : 510-486-5909 Web : CCI.LBL.gov

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43rd Mid-Atlantic Macromolecular Crystallography Meeting
by Charles Pemble 07 May '13

07 May '13

*43**rd** Mid-Atlantic Macromolecular Crystallography Meeting* *May 30 – June 1, 2013* *Duke University* *Durham, NC* This is a reminder for the 43rd Mid-Atlantic Macromolecular Crystallography Meeting, which will be held at Duke University in Durham, North Carolina. This meeting fosters discussion and interaction among structural biologists from around the Mid-Atlantic region. Registration and details can be found at http://www.mid-atlantic.org. The deadline for registration is May 15th. This year will feature keynote speaker Tom Terwilliger from Los Alamos National Lab, workshops on PHENIX and MolProbity, plus many talks & posters on new and exciting research. Presentations will be selected from submitted abstracts and preference will be given to young researchers. The best presentation and poster by a Graduate Student & Postdoctoral Fellow will be awarded (total of 4). We encourage you to submit your abstracts and sign up for the workshops as these are separate processes from registration. *DEADLINES:* Registration - May 15th Abstract submission for presentation - May 10th Abstract submission for poster - May 27th *IMPORTANT: *A block of rooms have been reserved at the Millennium hotel as well as the Duke dormitories. The deadline for reserving a room at the Millennium is May 9th. With best wishes on behalf of the primary Organizing Committee, Charlie Pemble _____________________________ Charles W. Pemble IV, Ph.D. Duke University Medical Center 308 Research Drive LSRC, A06 Durham, NC 27708 charles.pemble(a)duke.edu We thank our current sponsors for their generous contributions: Rigaku Americas Corporation, Agilent Technologies Inc., GlaxoSmithKline, Art Robbins, Bruker, HKL Research Inc., Molecular Dimensions, Hampton Research, Labcyte, GenScript, Genentech, Emerald BioSystems, TTP LabTech, Oxford Cryosystems, PhD Posters, Duke University School of Medicine, Duke Human Vaccine Institute, and Avanti Polar Lipids Inc.

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Morphing only a part of the model?
by Guenter Fritz 06 May '13

06 May '13

Dear all, I am working on a multisubunit protein complex (ca. 2000 residues). I would like to morph only a part or one subunit into the experimental electron density without morphing the rest of the protein. iIs there a way to select a subunit or a part of the model? Best regards, Guenter

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Find Difference peaks and holes
by John Rose 03 May '13

03 May '13

Hi, I can't seem to find the "Find Difference peaks and holes" utility in the PHENIX 1.8.2-1309 distribution. I found it to be very useful. Can you tell me where it is hiding. Regards, John John Rose Ph.D. Associate Professor B204B, The Fred C. Davison Life Sciences Complex 120 Green Street Department of Biochemistry and Molecular Biology University of Georgia Athens, GA 30602-7229 ========================================= Phone: 706-542-1750 Fax: 706-542-3077

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ReadySet & phenix.refine problem: NAG-Asn with hydrogen atoms
by Zhijie Li 29 Apr '13

29 Apr '13

Hello, I am refining my structure with hydrogen atoms on (phenix 1.8.2-1309). The protein contains two Asn-linked GlcNAc. I noticed two problems with respect to the NAG-Asn linkage in the presence of hydrogen atoms: 1) After running ReadySet (to add hydrogen to the PDB ab initio), the H1 on the NAG is pointing to the amino group of Asn, ie, the NAG seems to be interpreted as in its alpha anomer instead of beta (should be GlcNAc beta1,N-Asn). This is not really a problem as it doesn't seem to affect the refinement. 2) The H21 in Asn is correctly removed by ReadySet, leaving only H22. However, if I send this PDB to phenix.refine, the refinement will fail almost instantly complaining that H22 of the Asn has an unknown nonbonded energy type symbol. But if I simply change the atom name of H22 to H21, the refinement will proceed to completion with no problem, only leaving an H21 at the position where H22 should be. I was not sure what caused this error. So I trimmed my PDBs down to the Asn and NAG only, and fed the small PDB to ReadySet and phenix.refine (with an eff file indicating the NAG-Asn linkage) and got the same behavior: H22 on Asn - error; H21 on Asn - OK. I hope someone can look into this. Thanks in advance. Zhijie

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SIGF in refinement
by Aksyuk, Anastasia (NIH/NIAMS) [F] 29 Apr '13

29 Apr '13

Hi, I was wondering how SigFobs values are used in phenix refinement and how important those are. I am asking in view of P1 space group with low redundancy or even cryo-EM map based Fobs. Maybe you can point me to a useful reference on likelihood targets. I feel lost in those that I have. Thanks, --Anastasia ------------------------------ Anastasia A Aksyuk, PhD Laboratory of Structural Biology, NIAMS Bldg 50, Room 1511, 50 South Drive MSC 8025 N.I.H., Bethesda, MD 20892-8025 tel 301 451-8247 e-mail: anastasia.aksyuk(a)nih.gov

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Re: [phenixbb] clashscore discrepancies
by Alex Theodossis 27 Apr '13

27 Apr '13

Dear phenixbb, I have carried out refinement using the latest version of PHENIX, which gives improved molprobity stats. However, these stats still do not match those porvided by the molprobity server: PHENIX output Molprobity server Existing H-model Electron-cloud H-model All atom clashscore 2.57 4.2 (81st perc) 3.25 (88th percentile) Rotamer outliers 0.7 1.14 1.14 Overall score 1.04 1/24 (89th perc) 1.16 (93rd percentile) Given that PHENIX is now giving me the best results and all scenarios give reasonable results I am less inclined to be concerned. However, I still find these inconsistencies rather puzzling and am curious to learn what is causing them. Thanks in advance.

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phenix.elbow: Best ploy to generate restraints files for similar compounds ?
by Phil Jeffrey 25 Apr '13

25 Apr '13

I've got homologues of small molecule compounds I want to generate restraints for. The changes I'm looking at are converting O to S and moving a substituent around on a phenyl ring. I expect them to have similar conformations overall. Is the best method to: 1. create a new PDB file of the homolog, change atom types and do some primitive modeling, assuming that the --opt method will fix geometry snafus ~or~ 2. use a SMILES string, edit the numbering of the PDB file and regenerate the restraints from that Thanks Phil Jeffrey Princeton

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clashscore discrepancies
by Alex Theodossis 25 Apr '13

25 Apr '13

Dear phenixbb, Have refined a 1.05A structure with riding hydrogen model in phenix v1.8.1-1168 and doing final validation. The molprobity statistics output by phenix include: all atom clashscore = 4.12 Rotamer outliers = 0.71% Overall score = 1.2 Put through the molprobity server I am given the option of analysing with existing hydrogen model, or replacing with the 'electron-cloud parameters' they recommend for X-ray crystal structures. The results end up being very different: existing H model: electron-cloud model: All atom clash score: 4.66 (78th percentile) 3.72 (85th percentile) Poor rotamers: 1.14% 1.14% Overall score: 1.28 (88th percentile) 1.20 (92nd percentile) While all three scenarios give acceptable statistics, I am surprised by the inconsistency in the results given by molprobity, even when using the same hydrogen model. I am also wondering whether I am using a suboptimal riding model in my refinement (generated using phenix ready-set). Can anyone advise on the reason for these inconsistencies (is it just due to differences in the version of molprobity being run in each case). Is there anything I should be doing differently? Thanks in advance. On 18 November 2012 12:36, Alex Theodossis <alex.theodossis(a)monash.edu>wrote: > Hi, > > I'm attempting to install the latest version of phenix on mac 10.5.8, > using the command line installer. > > Installation proceeds well until towards the end, where it hangs on: > > precompiling .py files > > I can leave it for up to an hour without any further progress, but upon > typing: > > ctrl+c > > The installation comes back to life and proceeds to completion. > > Can anyone advise as to what might be causing this and on how reliable > this installation is going to be? > > Thanks in advance. > > >

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Postdoctoral positions in structure and function of membrane proteins at the NIH
by Anirban Banerjee 24 Apr '13

24 Apr '13

Dear all, Will you please bring this to the attention of suitable candidates ? Postdoctoral positions in structure and function of membrane proteins at the NIH The research group of Dr. Anirban Banerjee at the National Institutes of Health (NIH) is seeking candidates for postdoctoral fellows to start from September, 2013 or later. The broad interests of the lab are in membrane protein structure and function. We will combine macromolecular crystallography with functional assays with detergent-solubilized and liposome-reconstituted proteins to investigate the structural bases of the mechanisms of a number of membrane proteins. The lab is part of the Cell Biology and Metabolism (CBMP) program and is part of a large and diverse macromolecular crystallography and NMR community in the NIH campus. There is ample opportunity to interact with other scientific groups across a broad range of scientific disciplines. CBMP is part of the National Institute of Child Health and Human Development (NICHD). Our lab also has an affiliation to the National Institute of Neurological Disorders and Stroke (NINDS). The DC Metro area is a wonderful place to live, the weather is pleasant and the NIH is a vibrant international community. Candidates must be motivated and have a strong background in biochemistry and structural biology. Experience with protein expression, purification and crystallization is required. Prior experience in membrane protein biochemistry will be an added advantage. Candidates should hold a Ph.D. degree by the time they join and have no more than one year of postdoctoral experience at the time of applying. To apply, please send an email of interest to anirban.banerjee(a)nih.gov with a CV and a summary of previous research experience and future research interests. Best regards, Anirban

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install problem on Ubuntu 12.10
by Guan Hong Hsiang 24 Apr '13

24 Apr '13

Dear all: When I wanted to install phenix on my computer with ubuntu 12.10 (./install), the error message showed that "/usr/local/phenix-installer-1.8.2-1309/bin/install-from-binary:130:export:6 core : bad variable name. I am a newbie of phenix. Could anyone give me some suggestion? Thank you Hong-Hsiang

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phenix.refine - blocking sorry error
by Maxime Cuypers 24 Apr '13

24 Apr '13

hello, i just installed the latest buid of phenix : 1356 i now get the following error in phenix.refine - this was ok with previous version ' Sorry: Your refinement strategy includes anisotropic ADPs for hydrogen atoms refined individually; hydrogens should be left as isotropic even for individual refinement. ' which is the new specific command to add to the .params file that will force anisotropic refinement of deuteriums please? PS: this is needed for my neutron structure refinement. thanks for your help -- Dr. Maxime Cuypers Institut Laue Langevin (ILL), Deuteration lab Rue Jules Horowitz, 6, BP 156 38042 Grenoble cedex 9 France

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Clashes while searching second copy using phaser.AUTOMR
by Rojan Shrestha 22 Apr '13

22 Apr '13

Hello, I have same problem as in the following link which is the previous thread in this bb. If you solved the problem who posted, please let me know the solution how it can be tackled. http://www.phenix-online.org/pipermail/phenixbb/2009-April/013136.html I am trying to find 6 copies in ASU. The space group of diffraction set is P3121. I have obtained one copy with high TFZ scores (18.5) without clashes. However, while searching for second copy, Phaser gave so many clashes with no solution. As I read in above thread, it could be due to parallel pseudo-translation symmetry. Can somebody suggest me to overcome this type of problem? Regards, Rojan

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ICSG 2013 Abstract Deadline extended to May 15, 2013: International Conference on Structural Genomics - Structural Life Science
by Terwilliger, Thomas C 22 Apr '13

22 Apr '13

Dear Colleagues, We hope that you are planning to attend the International Conference on Structural Genomics 2013, which will be held in Sapporo, Hokkaido, Japan, July 29th – August 1st, 2013. ICSG2013-SLS is intended to provide an overview for the most recent developments in Structural Genomics and its impact on research in biology, medicine and disease, and to foster international collaboration among researchers. Five oral presentations will be chosen from the posters, and four poster prizes will be awarded (one HKL Research Best Technology Poster Prize and three Journal of Structural and Functional Genomics Poster Prizes)! Please submit your abstracts by May 15, 2013! You can see all the details of the conference at: http://www.c-linkage.co.jp/ICSG2013 The scientific topics covered in ICSG2013-SLS include the wider life science research fields with particular attention to drug discovery (small molecules and biopharmaceuticals), biotechnology and industrial issues while keeping strength in the high-throughput technologies and integration of hybrid methods. These technologies are now leading to the new field of “Structural Life Science”. In order to widen the opportunity to junior researchers to study more, we have organized several satellite workshops before ICSG2013- SLS (during the day on July 29, 2013). The topics will include “Small molecule screening”, “Automation of X-ray Structure Determination”, “Cell-free Protein Production”, “Automated NMR methods”, "Eukaryotic expression", “Interaction analyses" and "Bioinformatics”. We hope you also find the satellite workshops are informative and productive. The conference will be held in Keio Plaza Hotel Sapporo, in walking distance of Hokkaido University's main campus and Sapporo station. The summer in Sapporo is a great time to stay and enjoy the cool summer night of Japan. ICSG2-13-SLS is partially supported by the Grant-in-Aid for Scientific Research on Innovative Areas; “Structural Cell Biology”, “Intrinsically Disordered Protein” and “Transient Macromolecular Complexes”, from Ministry of Education, Culture, Sports, Science and Technology (MEXT) . We are looking forward to welcoming you to Sapporo this summer! Sincerely yours, Katsumi Maenaka, Ph.D. Chair, International Conference on Structural Genomics 2013 -Structural Life Science- (ICSG2013-SLS) Laboratory of Biomolecular Science and Center for Research and Education on Drug Discovery, Faculty of Pharmaceutical Sciences, Hokkaido University, Japan Soichi Wakatsuki Chair of Program Committee, ICSG2013-SLS Photon Science, SLAC National Accelerator Laboratory Department of Structural Biology School of Medicine Stanford University The International Structural Genomics Organization Executive Committee Shigeyuki Yokoyama Dino Moras Joel Sussman Jennifer Martin Aled Edwards Tom Terwilliger

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Refinement of perfect twined data of P31/P32
by miyatake 22 Apr '13

22 Apr '13

Hello, Phenix users, Now I'm triying to refine almost perfect twining data (mixing of P31 and P32). It has a twining operation of (h, k, l) = (-h, -k, l), detected by the xtriage program. The data qualigy seems to be good, deflacting to 1.6A resolution with Rmerge ~ 5%. When I refined the MR structures with phenix.refine, however, the model geometries have been worse as over the 20% residues fall into disallowed Ramachandran region, and Rfree stacked around 30%. I tried many options such as real space refinement, SA, bad rotamer fixation, NCS restrain,,.., but they seem to be not so effective as usual twininig cases.. One molecule consists of 202 residues, and three molecules are inolved in AU. Of course, I used the twin option (-h, -k, l) during the refinement. Manual collection of the models could not improve the model anymore. Would anyone please advise me how should I refine the twinned data? Sincerely, Hideyuki, M.

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Turn off non-bonded interactions to measure distance in case of unknown interaction
by Edward A. Berry 22 Apr '13

22 Apr '13

I have a phenix-specific question and a general chemistry question. In a ligand-bound structure, S in an aromatic ring (substituted thiophene) may be H-bonding NH1 of arginine guanidino group. The general question is whether thiophene S can act as H-bond acceptor (assuming one lone pair is in the aromatic pi system for a total of 6 e, the other lone pair is available as H-bond acceptor?). The phenix-specific question involves whether non-bonded interactions will push the atoms apart during refinement, and how to turn off the nonbonded interaction between these two atoms to allow the xray data to determine the position. Under geometry_restraints.edits I define a bond, but give a very large sigma of 1A. Will this effectively remove non-bonded interactions between these two atoms?

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dummy atom v.s. model with zero occupancy.
by Jinzhong Lin 21 Apr '13

21 Apr '13

Dear PHENIX users, In phenix, in order to improve bulk solvent model when the model is not complete, you can add dummy atom and set "refinement.mask.ignore_zero_occupancy_atoms=False". My question is that can I just put a model ( I know the approximate position for the model) there instead of dummy atoms and set all atoms in the model zero occupancy and keep the same setting. Will I get the same result? I suspect that model will still be used to calculate the Fc even though the occupancy is zero, so the map will be severely model biased. Lithlin

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Re: [phenixbb] how to fix a side chain not to be refined?
by Rongjin 19 Apr '13

19 Apr '13

Hi Nat, You are right. By turning off the "fix rotamers" the Arg does not move any more. Best Rongjin ------------------ Original ------------------ From: "Nathaniel Echols"<nechols(a)lbl.gov>; Date: Apr 20, 2013 To: "PHENIX user mailing list"<phenixbb(a)phenix-online.org>; Subject: Re: [phenixbb] how to fix a side chain not to be refined? On Fri, Apr 19, 2013 at 2:14 PM, Rongjin <guanrj(a)foxmail.com> wrote: > I have an Arg with flexible side chain, close to an iodide ion. Every time > I refine it, the side chain of Arg will push the iodide ion away, even I > selected The syntax below looks correct to me, although you should probably do the same thing for the ADP selection, but it's a little disturbing that it's doing this. Do you have rotamer correction turned on? That may be the problem, if it is trying to fit to the difference density (I don't think it's very aware of nearby atoms). -Nat > the Arg not to be refined: > > refine { > strategy = *individual_sites individual_sites_real_space rigid_body \ > *individual_adp group_adp tls *occupancies group_anomalous > sites { > individual = "not (chain A and resid 40)" > } > } > > > The residue 40 in Chain A is that Arg close to the Iodide. Is this selection > correct? > How to fix the side chain of Arg in refinement? > > I attached two pictures, one before and one after refinement, the local area > of the Arg > and the iodide. The iodide is bound to an Asn residue of one symmetric > molecule. > > Thank you > Rongjin Guan > > > > _______________________________________________ > phenixbb mailing list > phenixbb(a)phenix-online.org > http://phenix-online.org/mailman/listinfo/phenixbb > _______________________________________________ phenixbb mailing list phenixbb(a)phenix-online.org http://phenix-online.org/mailman/listinfo/phenixbb .

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how to fix a side chain not to be refined?
by Rongjin 19 Apr '13

19 Apr '13

Hello I have an Arg with flexible side chain, close to an iodide ion. Every time I refine it, the side chain of Arg will push the iodide ion away, even I selected the Arg not to be refined: refine { strategy = *individual_sites individual_sites_real_space rigid_body \ *individual_adp group_adp tls *occupancies group_anomalous sites { individual = "not (chain A and resid 40)" } } The residue 40 in Chain A is that Arg close to the Iodide. Is this selection correct? How to fix the side chain of Arg in refinement? I attached two pictures, one before and one after refinement, the local area of the Arg and the iodide. The iodide is bound to an Asn residue of one symmetric molecule. Thank you Rongjin Guan

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Unusually low B factors
by Brennan Bonnet 17 Apr '13

17 Apr '13

Hi all, I have produced xenon derivatives for SAD phasing of several standard proteins by placing them under xenon gas pressure and then immediately flash freezing in liquid nitrogen. Everything has worked out nicely as far as successful phasing, nice models, and low R factors. Now I've noticed that the B values have decreased from initial processing values. Several rounds of manual changes and phenix.refine have caused B values to decrease each round of refinement to values of around 10 (I guess 20 is a more common value). Does anyone know why this is happening or what the explanation might be? I'm not sure what other information to include so please contact to discuss further. Thanks! ~Brennan

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Exclude atoms from refinements
by Simon Kolstoe 16 Apr '13

16 Apr '13

Hi, I want to exclude some atoms from refinement altogether in phenix.refine. Googling this I came up with the following link: http://phenix-online.org/pipermail/phenixbb/2011-December/018020.html however was wondering how you implement this sort of thing in the GUI? Thanks, Simon --------------------------------------------------------------- Dr. Simon Kolstoe Honorary Senior Research Associate Laboratory for Protein Crystallography Wolfson Drug Discovery Unit University College London Rowland Hill Street, London, NW3 2PF http://www.ucl.ac.uk/~rmhasek ---------------------------------------------------------------

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Pause phenix runs
by Mark J van Raaij 12 Apr '13

12 Apr '13

Just wondering if being able to pause a phenix run would be something useful. Currently, it appears one can only abort. Currently doing refinement of a large structure, and doing several simultaneous runs with different options is slowing down the computer too much. Individual runs take about 72 hrs, so o/n is not an option (o/weekend will be...) (or is it possible and I have missed it?) if the FPLC can do it, why not phenix? Seriously, I don't know how easy or difficult it would be to implement, or if paused runs would still take up too many resources, but it seems like something that might be useful - i.e. set several long runs running, and temporarily interrupt them for other jobs on the same computer (like watching youtube movies, as some people seem to think I do all day...) Mark J van Raaij Lab 20B Dpto de Estructura de Macromoleculas Centro Nacional de Biotecnologia - CSIC c/Darwin 3 E-28049 Madrid, Spain tel. (+34) 91 585 4616 http://www.cnb.csic.es/~mjvanraaij

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