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Re: [phenixbb] [ccp4bb] Tantalum anomalous signal of a low-resolution SAD dataset with lattice-translocation disorder
by Christian Roth 21 Oct '15

21 Oct '15

Hi, difficult to say why it doesn't work. Generally as mentioned already automatic model buildíng will most likely fail at this resolution. However why do you think your sites are wrong? Ta-cluster are regular compounds and the sites should fit with the geometry of the cluster at least. Anyway, molecular replacement using anomalous differences and the cluster as search model might work in that case as well, if direct experimental phasing fails. Cheers Christian Am 21.10.2015 um 05:19 schrieb Shun Liu: > Dear CCP4 colleagues, > > We are working on a Ta6Br14 cluster-SAD dataset (4.3 angstroms) with lattice-translocation disorder (with a total Rmerge of 0.16). Both SHELXC and Xtriage gave the similar positive result about the anomalous signal (See below). > > ------------------------------------------------------------------------------------------------- > Resl. Inf. 9.58 7.61 6.65 6.04 5.61 5.27 5.01 4.79 4.61 4.45 4.31 > N(data) 1427 1410 1410 1409 1385 1452 1367 1399 1367 1395 1385 > <I/sig> 76.3 52.1 26.7 13.7 8.7 6.1 5.1 5.0 3.9 2.7 1.8 > %Complete 98.8 99.9 100.0 99.9 100.0 100.0 100.0 99.9 99.7 99.6 95.8 > <d"/sig> 11.67 9.67 6.12 3.73 2.50 1.59 1.21 0.99 0.84 0.79 0.68 > ------------------------------------------------------------------------------------------------ > unused: - 49.7669 [ 0/29 ] > bin 1: 49.7669 - 9.2629 [3086/3100] 0.8212 > bin 2: 9.2629 - 7.3599 [3031/3031] 0.7689 > bin 3: 7.3599 - 6.4318 [3073/3073] 0.5380 > bin 4: 6.4318 - 5.8447 [3071/3075] 0.2933 > bin 5: 5.8447 - 5.4263 [3054/3054] 0.1230 > bin 6: 5.4263 - 5.1067 [3070/3071] 0.0363 > bin 7: 5.1067 - 4.8512 [3038/3039] 0.0126 > bin 8: 4.8512 - 4.6402 [3028/3048] 0.0074 > bin 9: 4.6402 - 4.4617 [2955/3037] 0.0021 > bin 10: 4.4617 - 4.3078 [2730/3059] 0.0000 > unused: 4.3078 - [ 0/0 ] > -------------------------------------------------------------------------------------------------- > The Tantalum sites and the initial map seem ambiguous, with which automatic model-building failed. > I am wondering whether the lattice-translocation disorder of the dataset impacts the reliability of the anomalous signal, tantalum sites and the initial map. If it does, how can we decrease its impact? If it doesn't, is it possible to find the accurate Ta sites and generate an interpretable map suitable for model-building with this dataset? (After all, it has been reported that Ta sites can be found at 6A resolution.) > > Any suggestion and comment will be highly appreciated! > > Best, > Shun >

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Tantalum anomalous signal of a low-resolution SAD dataset with lattice-translocation disorder
by Shun Liu 21 Oct '15

21 Oct '15

Dear Phenix colleagues, We are working on a Ta6Br14 cluster-SAD dataset (4.3 angstroms) with lattice-translocation disorder (with a total Rmerge of 0.16). Both SHELXC and Xtriage gave the similar positive result about the anomalous signal (See below). ------------------------------------------------------------------------------------------------- Resl. Inf. 9.58 7.61 6.65 6.04 5.61 5.27 5.01 4.79 4.61 4.45 4.31 N(data) 1427 1410 1410 1409 1385 1452 1367 1399 1367 1395 1385 <I/sig> 76.3 52.1 26.7 13.7 8.7 6.1 5.1 5.0 3.9 2.7 1.8 %Complete 98.8 99.9 100.0 99.9 100.0 100.0 100.0 99.9 99.7 99.6 95.8 <d"/sig> 11.67 9.67 6.12 3.73 2.50 1.59 1.21 0.99 0.84 0.79 0.68 ------------------------------------------------------------------------------------------------ unused: - 49.7669 [ 0/29 ] bin 1: 49.7669 - 9.2629 [3086/3100] 0.8212 bin 2: 9.2629 - 7.3599 [3031/3031] 0.7689 bin 3: 7.3599 - 6.4318 [3073/3073] 0.5380 bin 4: 6.4318 - 5.8447 [3071/3075] 0.2933 bin 5: 5.8447 - 5.4263 [3054/3054] 0.1230 bin 6: 5.4263 - 5.1067 [3070/3071] 0.0363 bin 7: 5.1067 - 4.8512 [3038/3039] 0.0126 bin 8: 4.8512 - 4.6402 [3028/3048] 0.0074 bin 9: 4.6402 - 4.4617 [2955/3037] 0.0021 bin 10: 4.4617 - 4.3078 [2730/3059] 0.0000 unused: 4.3078 - [ 0/0 ] -------------------------------------------------------------------------------------------------- However, the tantalum sites we found and the initial map seem ambiguous, with which automatic model-building failed. I am wondering whether the lattice-translocation disorder of the dataset impacts the reliability of the anomalous signal, tantalum sites and the initial map. If it does, how can we decrease its impact? If it doesn't, is it possible to find the accurate Ta sites and generate an interpretable map suitable for model-building with this dataset? (After all, it has been reported that Ta sites can be found at 6A resolution.) Any suggestion and comment will be highly appreciated! Best, Shun

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Queueing system commands for parallel Phenix/Rosetta jobs
by Yonglin Hu 21 Oct '15

21 Oct '15

Hello All, We have setup a cluster with several executing nodes and we would like to run our Phenix/Rosetta molecular replacement jobs on the cluster. In the Job Control screen, I put "qsub -q all.q -o $HOME/log1 -e $HOME/log1" as command to use to run multiple jobs. But I am not sure what I should use for "queueing system commands". We are using SGE as the queueing system on our cluster. Any help would be greatly appreciated! Yonglin Hu

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on cif file
by Smith Lee 19 Oct '15

19 Oct '15

Dear All, My protein containing the ADP-AlF3-Mg. In the protein ADP-AlF3-Mg should be very close to each other so that they look like the ATP binding with the Mg. I have prepared the ADP.cif and the ALF3.cif, and have merged the 2 cif files with elbow.join_cif_files. However when I run phenix.real_space_refine with the merged cif file, there was a tendency that ADP, ALF3 and Mg separated from each other based on the map density, which seems did not comply with the prior knowledge on that ADP-AlF3-Mg should occupies the space of ATP-MG. Will you please advice how to have ADP-AlF3-Mg occupies the space of ATP-MG rather than separated by the refine? Best regards. Smith

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Re: [phenixbb] How to convert XDS_ASCII.HKL to MTZ
by Whitley, Matthew J 19 Oct '15

19 Oct '15

Hi Murpholino, I’m not sure if your Phenix commands are correct, but you might try using the program that comes natively with XDS for converting between file formats. That program is XDSCONV. (As a side note, I think that some of the XDSCONV functions require CCP4 to be installed, so if you don’t have CCP4 installed you might need to stick to converting with Phenix.) You can find step by step instructions for generating an MTZ file from XDS output under the “Typical Use” heading here: http://strucbio.biologie.uni-konstanz.de/xdswiki/index.php/Xdsconv More info about XDSCONV: http://homes.mpimf-heidelberg.mpg.de/~kabsch/xds/html_doc/xdsconv_parameter… Good luck! Matthew On Oct 19, 2015, at 12:16 PM, <phenixbb-request(a)phenix-online.org> <phenixbb-request(a)phenix-online.org> wrote: > > Message: 5 > Date: Mon, 19 Oct 2015 11:16:17 -0500 > From: Murpholino Peligro <murpholinox(a)gmail.com> > To: PHENIX user mailing list <phenixbb(a)phenix-online.org> > Subject: [phenixbb] How to convert XDS_ASCII.HKL to MTZ > Message-ID: > <CAMdBu0Uejh0k+Wih1KaF+c=FRJ3fKg+LhQrfuGhUs1tJ8x8HwQ(a)mail.gmail.com> > Content-Type: text/plain; charset="utf-8" > > Just want to be sure. > I want to generate the mtz file for refinement. The last step of XDS leaves > me with intensities and space group 89. So I need > > phenix.reflection_file_converter --generate-r-free-flags > --write-mtz-amplitudes --mtz_root_label=FOBS --mtz=d01.mtz > --change-to-space-group=96 XDS_ASCII.HKL > > Is this right? > > Thanks again >

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How to convert XDS_ASCII.HKL to MTZ
by Murpholino Peligro 19 Oct '15

19 Oct '15

Just want to be sure. I want to generate the mtz file for refinement. The last step of XDS leaves me with intensities and space group 89. So I need phenix.reflection_file_converter --generate-r-free-flags --write-mtz-amplitudes --mtz_root_label=FOBS --mtz=d01.mtz --change-to-space-group=96 XDS_ASCII.HKL Is this right? Thanks again

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refinement of coordinated ions
by Irina PROKHOROVA 18 Oct '15

18 Oct '15

Hello, I have many Osmium ions in my structure. They have 6 coordinated ammonium groups and I would like to refine the B-factors for them: one B-factor for Os in each residue and one B-factor for all 6 ammonium groups in each residue. For this I have chosen *group_adp with following syntax adp { group_adp_refinement_mode = one_adp_group_per_residue \ two_adp_groups_per_residue *group_selection group = chain O and element Os group = chain O and element N } I wanted to refine only B-factor for Osmiums in this refinement. To avoid the bias from previously refined values of B-factor I presetted B-factors for all Osmiums to 50. As a result I got the change of B-factors for all residues in the model to the same value (which probably means that all model was refined as one group) and R-free increased from 0.25 to 0.34. What should be the correct syntax for this refinement? Should I also refine ions coordinates for a good refinement of B-factors? Irina

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Fw: on phenix.real_space_refine
by Smith Lee 18 Oct '15

18 Oct '15

Dear Pavel, On Oct 9, 2015 I have e-mailed to you my experience with 1.9-1692 and version 1.10 (and 1.10.1) on phenix.real_space_refine. Today I just completed another protein phenix.real_space_refine. For the following comparison runs, the initial model same, the map same, all 5 macro_cycles default run. For version 1.10.1, the results were as following, Map CC (whole unit cell)： 0.672Map CC （around atoms）： 0.761rmsd （bonds): 0.01rmsd (angles): 1.55All-atom clahscore: 39.27Ramachandran outlier: 0.98%Ramachandran allowed: 12.71%Ramachandran favoured: 86.31%Rotamer outliers: 0.91%C-beta deviations: 0 For version 1.9-1692, the results were as following, Map CC (whole unit cell)： 0.645Map CC （around atoms）： 0.687rmsd （bonds): 0.0071rmsd (angles): 1.15All-atom clahscore:16.88Ramachandran outlier: 2.00%Ramachandran allowed: 6.23%Ramachandran favoured: 91.77%Rotamer outliers: 0.05%C-beta deviations: 0 For the version 1.10.1 phenix.real_space_refine, the most unacceptable was on the clashscore, the whole refine process seems did not care on the clashcore refine at all. I am looking forward to getting an explanation from you on the causes of the difference for phenix.real_space_refine for the 2 versions, and the possible strategy we can take to eliminate the difference to get the acceptable results. Best regards. Smith On Friday, October 9, 2015 12:52 AM, Smith Lee <smith_lee123(a)yahoo.com> wrote: Dear Pavel， As I have told you, I have tried phenix.real_space_refine for 2 protein with version 1.9-1692 and version 1.10 (and 1.10.1) for many times, with cif or without cif, with or without annealing. The general feedback was that the version 1.9-1692 was much much better for the final analysis of the PDB given at the end of the run of the phenix.real_space_refine. Especially for clashing score, version 1.9-1692 can gave acceptable results (below 20 or around 20), but the clashing score given by the above 2 new versions of phenix.real_space_refine was very high (around 50 or 100). For C-beta deviations, the output by 1.9-1692 was also much better Before I have tried the other nightly built versions of phenix.real_space_ refine between version 1.9-1692 and version 1.10 (and 1.10.1) , all I gave acceptable output analysis (something like the Molprobity analysis at the end of the run). Thus may I ask the reason you have version 1.10 (and 1.10.1) phenix.real_space_refine was because you have realized that the result given by version 1.9-1692 was too satisfactory? One direct concern of me was that you have recommended not to use the old version phenix. But I cannot get the satisfactory result with the new version phenix.real_space_refine. The other concern of me was that, I have noticed there were publications In Nature and Science which wrote they use phenix.real_space_refine to got the results in Nature and Science. The Phenix they used should be the versions earlier than version 1.10. Considering what I mentioned above, the Nature and Science results may be not so correct, if new version of phenix was considered as much better than the old version. Thus am I the only user who communicated with you on the high clashing score given by the new version of phenix？Can you give some comments on what I mentioned above on the high clashing score given by the new version phenix.real_spac_refine? Best regards. Smith

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Visualise NCS operators
by Morris, Kyle 16 Oct '15

16 Oct '15

Dear Phenix community, After running $ phenix.find_ncs what would be the best way to visualise the operators found and detailed in the *.ncs_spec file superimposed on the map used in the search? Thanks for your help! Best wishes, Kyle

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Postdoctoral Position in Structural Biology at ICR, London, UK
by Sebastian Guettler 16 Oct '15

16 Oct '15

Dear PHENIX Community, Please see below for details of a postdoctoral position in our team. Best wishes, Sebastian Guettler Postdoctoral Position in Protein Biochemistry / Structural Biology / Electron Microscopy We seek to appoint a Postdoctoral Training Fellow to the research team led by Dr Sebastian Guettler in the Divisions of Structural and Cancer Biology at The Institute of Cancer Research in London, UK. The position is funded by Cancer Research UK. The successful applicant will undertake biochemical, structural and functional studies on members of the ADP-ribosyltransferase (or “PARP”) family of proteins and macromolecular complexes containing PARPs. Our goal is to gain insight into the control and cellular roles of these enzymes with a specific interest in ADP-ribosyltransferase-dependent signalling processes linked to cancer. Applicants should possess a PhD in a biological science such as biochemistry or molecular biology. Experience with recombinant DNA techniques, high-quality protein expression and purification, protein characterisation, structural biology, as well as some experience in structural electron microscopy are essential. Appointment will be on a Fixed Term Contract for 3 years in the first instance. The full salary scale is in the range from £29,517 to £42,187 p.a. inclusive. Starting salary will be based on previous postdoctoral experience. To apply, please submit your CV and covering letter online. Please include the names and addresses of two referees. For further details and instructions on how to apply, please refer to our online vacancy and recruitment site at http://www.icr.ac.uk/jobsearch, quoting job reference number 1494999 for this position. While applications need to be placed via our online system, informal inquiries can be sent to Dr Sebastian Guettler at sebastian.guettler(a)icr.ac.uk<mailto:[email protected]>. Please do not submit your formal application by email! Summary: Closing Date of vacancy: 31 Oct 2015 Type of Contract: Fixed Term, Postdoctoral Training Fellow Length of Contract: 3 years Salary Range: £29,517 p.a - £42,187 p.a. inclusive (full salary scale) Work Location: London Research in the Divisions of Structural and Cancer Biology focuses on the functional, biochemical, and structural characterisation of cellular processes relevant to cancer and cancer therapy. The Division of Structural Biology has managed state-of-the-art facilities for protein crystallography (crystallisation robots, Bruker Microstar and CCD detector), cryo-electron microscopy (FEI Tecnai F20 and T12), and protein production with expertise in multi-subunit expression (insect cell, yeast and bacterial expression, including a 60-litre fermentor). The Division is also well-equipped with instrumentation for biophysical analysis (e.g. ITC, fluorescence, multi-angle light scattering). The Division of Cancer Biology provides a state-of-the-art infrastructure for mammalian cell culture, imaging, mammalian genetics, chemical biology and proteomics. The Institute of Cancer Research: Royal Cancer Hospital, a charitable Company Limited by Guarantee, Registered in England under Company No. 534147 with its Registered Office at 123 Old Brompton Road, London SW7 3RP. This e-mail message is confidential and for use by the addressee only. If the message is received by anyone other than the addressee, please return the message to the sender by replying to it and then delete the message from your computer and network.

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