- phenixbb - phenix-online.org

reminder - postdoc position at Astex
by Gabor Bunkoczi 23 Feb '21

23 Feb '21

Dear All, Just a quick reminder that Astex has an opening for a skilled post-doctoral researcher with an interest in software development. The aim of the project is to develop new methods for automated ligand fitting guided by experimental electron density/electrostatic potential maps, and will focus on locating, sharpening and classifying unmodelled regions of density, determining the identity of the binder using density shape and other physical properties derived from the surroundings, and subsequently fitting the molecule taking potential interactions with the environment into account. For more information about the position, please see the full advert: https://astx.com/wp-content/uploads/2021/01/SI-Post-Doc-2019-Mol-Sci-new-ad… Interested candidates should liaise with the HR department. For informal enquiries, please feel free to contact me directly. Best wishes, Gabor Bunkoczi Astex Therapeutics Ltd 436 Cambridge Science Park Cambridge CB4 0QA, UK This email and any attachments thereto may contain private, confidential, and privileged material for the sole use of the intended recipient. Any review, copying or distribution of this email (or any attachments thereto) by others is strictly prohibited. If you are not the intended recipient, please delete the original and any copies of this email and any attachments thereto and notify the sender immediately.

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Virtual Annual CCP4/APS Crystallographic School (2021) in the US
by Xu, Qingping 22 Feb '21

22 Feb '21

Dear Colleagues, We are pleased to announce the virtual annual CCP4/APS crystallographic school "From data collection to structure refinement and beyond" will be held on June 14-25, 2021 at the Advanced Photon Source (APS), Argonne National Laboratory (ANL), near Chicago, Illinois, USA. All details can be found at the school website: http://www.ccp4.ac.uk/schools/APS-2021/index.php. Dates: June 14 through 25, 2021 (with a weekend break on June 19 and 20th) Location: Virtual/Remote The school comprises two parts: remote data collection workshop and virtual crystallographic computing workshop. Data collection workshop includes beamline training, data collection on GM/CA at APS beamlines 23ID-D and 23ID-B equipped with Pilatus3 6M and Eiger 16M detectors respectively, and data processing. For data collection, only the participants' crystals will be used. Crystallographic computation workshop will feature many modern crystallographic software packages taught by authors and other experts. The daily schedule will be organized in three sections - lectures, tutorials, and hands-on (interactive troubleshooting of the technical difficulties the participants face in their projects). We have had considerable success resolving these problems in past years, attested by resulting publications (see http://www.ccp4.ac.uk/schools/APS-school/publications.php). A sample program, contact info and other details can be found at the School website. Applicants: Graduate students, postdoctoral researchers and early-career faculty, along with commercial/industrial researchers are encouraged to apply. Only about 20 applicants will be selected for participation. Participants of the workshop are strongly encouraged to bring their own problem data sets or crystals so the problems can be addressed during data collection and/or computation workshops. Application: Application deadline is April 1st, 2021. To apply, visit https://www.ccp4.ac.uk/schools/APS-2021/application.php. Fees: There is no fee for attending the virtual school for the chosen applicants. We hope to see you at the school. Charles, Garib and Qingping

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Re: [phenixbb] Autosol help: FOM is great but map is trash
by Kay Diederichs 19 Feb '21

19 Feb '21

Hi Alex, I'm not a Phenix developer, but I have seen many cases like this, where reflections can be indexed with different cells, and one unit cell axis differs by a factor of 2. This just means that every second reflection (namely the odd one) along the reciprocal space direction corresponding to that axis is weak. It is not a priori clear which of the two choices is the best for structure solution and refinement. This depends on whether the weak reflections actually carry structural information (e.g. about tNCS with a translation close to 0.5), or whether they are a result of a crystal defect. It may sometimes happen that the crystal form is not suitable at all, since both choices don't refine well! Concerning P212121, P21212, P2122, etc: these are all primitive orthorhombic, and data processing alone cannot tell you which one is correct - they give identical data processing statistics. Typically, absent reflections along h00 0k0 00l should tell about screw axes, but in case of tNCS and two different cell choices the correct choice may not be obvious. Molecular replacement and subsequent refinement of all possibilities should help. Furthermore, contrary to what you write, it _is_ possible for all the orthorhombic space groups to account for reflections in both cell choices, so don't jump to conclusions too early. Just explicitly specify the (big or small) cell parameters in the data processing program of your choice! It may also help to try a different data processing package. Hope this helps, Kay Am 19.02.2021 um 04:28 schrieb phenixbb-request(a)phenix-online.org: > Message: 4 > Date: Thu, 18 Feb 2021 18:15:15 -0500 > From: Alex Johnson<algejohnson(a)gmail.com> > To:[email protected] > Subject: [phenixbb] Autosol help: FOM is great but map is trash > Message-ID:<6BE0A20E-F46B-482A-A675-22B685A2AFBE(a)gmail.com> > Content-Type: text/plain; charset="utf-8" > > Hi Phenix team, > > I am attempting to phase a protein structure from SeMet-labeled crystals and have encountered some issues in using Autosol that appear unusual, as they have not been encountered by any members of my group. The short summary is that while I achieved great statistics from beam line collection and Autosol produces solutions with FOM (>0.4) from multiple crystals processed to ~2.4-3.0 A, the output maps are of poor quality and Autobuild can never build with Rfree <0.5. There is only one methionine for the ~30 kDa protein, but the crystals appear to have good anomalous signal with anomalous multiplicity >15 overall and delanon in the inner shell >0.5. There appears to be translational NCS and the dataset can be processed with good statistics in several space groups (P212121, P21212, P2122, etc) but this doesn?t solve the phase problem. The axis drops by half in processing in the P212121 versus P21212 space groups: from one crystal they are 48.37 101.56 110.39 90 90 90 (in P212121) and 101 > .62 48.35 55.19 90 90 90 (in P21212). > > If anyone in the community has encountered similar problems please let me know! I am all ears to any possible solutions and am also installing additional methionine for labeling but unsure whether this will overcome the other issues. > > Thanks, > > Alex Johnson > Postdoc, Kranzusch lab, Dana-Farber Cancer Institute > algejohnson(a)gmail.com <mailto:[email protected]> > > > > > >

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Scientist Antibody Production at the Institute for Protein Innovation in Boston, USA
by Christopher Bahl 19 Feb '21

19 Feb '21

Scientist Antibody Production Reports to the Head of Biological Discovery, Dr. Rob Meijers The Institute for Protein Innovation (IPI) is a nonprofit research organization advancing protein science to accelerate research and improve human health. Located on the Harvard Medical School campus in the Boston Longwood Medical Area, IPI is pioneering a new way of doing science that uniquely combines the freedom of academia with the high-throughput scaling of industry to take on transformative projects. IPI has built a robust platform and a repository of synthetic antibodies and other protein tools to share with the biomedical community through licensing and collaboration. The Institute’s deep protein expertise, collaborative spirit and fit-for-purpose antibodies are powering new biomedical and therapeutic discoveries for academic and industrial researchers worldwide. Job Overview IPI’s flagship project, the “Antibody Initiative,” aims to create well-characterized, synthetic antibodies against human cell surface receptors and their ligands, particularly those that are traditionally difficult to target. These antibodies may have high therapeutic and diagnostic potential but will also serve to enable basic biological discovery. The Scientist, Antibody Production will play an essential role in the high-throughput production, characterization and formulation of synthetic antibodies for this Initiative and other key IPI projects. Specifically, the successful candidate will join the Protein Group, a dynamic team generating high-quality antibodies and developing new technologies for conjugation. Having completed a doctorate in protein chemistry, the successful candidate will bring to the job experience in mammalian expression, purification and reagent-grade preparation of glycoproteins. The candidate will be adept in quality control and process management and reliably provide detailed documentation on experiments completed. The Scientist will work in close collaboration with the other Antibody Initiative team members and enjoy the opportunity to contribute to a wide array of novel biological discovery projects. As this position is highly collaborative, strong communication skills are needed. The Scientist will engage with outside collaborators, translate their biological questions into novel protein reagents and share these reagents with the larger research community through abstracts, presentations and publications. Responsibilities and Duties -Express, purify, and characterize glycoproteins from mammalian cells -Leverage robotics and automation to increase the quality and throughput of antibody production at IPI -Formulate and package antibodies for use by collaborators and customers -Supervise research associates and coordinate with other scientists to ensure efficient, high-quality antibody production -Actively lead the experimental design and data production for collaborations and scientific publications -Participate in meetings and conferences to engage and recruit outside research groups to participate in the Antibody Initiative Qualifications -PhD degree in mammalian protein expression, protein biochemistry, or structural biology -Well acquainted with best practices in cell culture and reagent production -Ability to work in a fast-paced, intellectually challenging environment -Enthusiastic team member with a strong commitment to mission and a desire to conduct impactful science -Good organizational and documentation skills -The ideal candidate will have recently completed a PhD and have a desire to explore a more entrepreneurial position IPI is an independent 501(3)(c) nonprofit research organization and an equal opportunity employer. The Institute celebrates diversity and is committed to creating an inclusive environment for all employees. If interested, please email your resume to . -- PLEASE NOTE: This message, including any attachments, may include privileged, confidential and/or inside information belonging to IPI. Any distribution or use of this communication by anyone other than the intended recipient(s) is strictly prohibited and may be unlawful. If you are not the intended recipient, please notify the sender by replying to this message and then delete it from your system. Thank you.

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Autosol help: FOM is great but map is trash
by Alex Johnson 18 Feb '21

18 Feb '21

Hi Phenix team, I am attempting to phase a protein structure from SeMet-labeled crystals and have encountered some issues in using Autosol that appear unusual, as they have not been encountered by any members of my group. The short summary is that while I achieved great statistics from beam line collection and Autosol produces solutions with FOM (>0.4) from multiple crystals processed to ~2.4-3.0 A, the output maps are of poor quality and Autobuild can never build with Rfree <0.5. There is only one methionine for the ~30 kDa protein, but the crystals appear to have good anomalous signal with anomalous multiplicity >15 overall and delanon in the inner shell >0.5. There appears to be translational NCS and the dataset can be processed with good statistics in several space groups (P212121, P21212, P2122, etc) but this doesn’t solve the phase problem. The axis drops by half in processing in the P212121 versus P21212 space groups: from one crystal they are 48.37 101.56 110.39 90 90 90 (in P212121) and 101.62 48.35 55.19 90 90 90 (in P21212). If anyone in the community has encountered similar problems please let me know! I am all ears to any possible solutions and am also installing additional methionine for labeling but unsure whether this will overcome the other issues. Thanks, Alex Johnson Postdoc, Kranzusch lab, Dana-Farber Cancer Institute algejohnson(a)gmail.com <mailto:[email protected]>

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Post-Doctoral Position at the University of Illinois to Study Ribosome Structure
by Yury Polikanov 16 Feb '21

16 Feb '21

*Post-Doctoral Position in Structural Biology at the University of Illinois at Chicago (UIC) to Study the Structure and Function of the Ribosome* The Polikanov Laboratory at the University of Illinois at Chicago (IL, USA) is looking for a talented, skillful, and highly motivated researcher to fill the available *NIH-funded Post-Doctoral Scholar *position to study the structure and function of the bacterial ribosome. The research in the Polikanov Laboratory is focused on understanding the basic principles of protein synthesis in bacteria, the modes of action of ribosome-targeting antibiotics, and the mechanisms of drug resistance at a structural level. More information about Dr. Polikanov research can be obtained from the most recent publications (*https://www.ncbi.nlm.nih.gov/sites/myncbi/yury.polikanov.1/bibliography/50395623/public/?sort=date&direction=descending <https://www.ncbi.nlm.nih.gov/sites/myncbi/yury.polikanov.1/bibliography/503…>*) or found on the web-page (https://bios.uic.edu/bios/people/faculty/yury- polikanov). The successful candidate will be involved in the determination of high-resolution structures of ribosome functional complexes with various protein translation factors, tRNAs, and antibiotics using macromolecular X-ray crystallography and potentially cryo-EM techniques. *Basic qualifications:* - PhD in chemistry, biochemistry , biophysics, or a related field; - Practical skills with the most standard methods currently used in molecular biology, and biochemistry (such as protein expression, purification); - Experience in structural biology (X-ray crystallography or cryo-EM); - Experience in the RNA biology field is not required but highly desired; *The successful candidate is also expected to:* - Act independently in terms of planning and executing the experiments; - Compile, analyze, and prepare data for publications; - Assist with training of graduate and undergraduate students. Review of applications will begin immediately and will continue until the position is filled. Interested candidates should apply by 04/16/2021 using the following link: https://jobs.uic.edu/job-board/job-details?jobID=142760 or email directly to Dr. Polikanov - yuryp(a)uic.edu. Please make sure to include CV, cover letter, and contact information for three references. *For full consideration, in **the cover letter, describe** how your previous research accomplishments and scientific interests fit with the scope of the research in the Polikanov research group.* *The University of Illinois at Chicago is an Equal Opportunity, Affirmative Action employer. Minorities, women, veterans, and individuals with disabilities are encouraged to apply.* *Offers of employment by the University of Illinois may be subject to approval by the University’s Board of Trustees and are made contingent upon the candidate’s successful completion of any criminal background checks and other pre-employment assessments that may be required for the position being offered. Additional information regarding such pre-employment checks and assessments may be provided as applicable during the hiring process.* *The University of Illinois System requires candidates selected for hire to disclose any documented finding of sexual misconduct or sexual harassment and to authorize inquiries to current and former employers regarding findings of sexual misconduct or sexual harassment. For more information, visit **https://www.hr.uillinois.edu/cms/One.aspx?portalId=4292&pageId=1411899 <https://www.hr.uillinois.edu/cms/One.aspx?portalId=4292&pageId=1411899>*

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postdoctoral position in cryo-EM at LMB in Jan Löwe's group
by Jan Löwe 16 Feb '21

16 Feb '21

Dear all, My group has a grant-funded job opening for a postdoctoral structural biologist to work on problems in archaeal and bacterial cell biology. The aim is to understand cell division and vesicle formation at the molecular or even atomic scales. The work will involve state-of-the-art cryo-EM, biochemistry and tomography (and who knows, maybe even crystallography and/or NMR). We are surrounded here at LMB by many experts and a lot of exciting instrumentation, and given the current state of the project, only the sky (and our imagination) is the limit. Please find the job advert here: https://www.nature.com/naturecareers/job/postdoctoral-scientist-structural-… <https://www.nature.com/naturecareers/job/postdoctoral-scientist-structural-…> Please do get in touch if you would like to know more before applying. Best wishes, Jan Jan Löwe MRC Laboratory of Molecular Biology Francis Crick Avenue Cambridge Biomedical Campus Cambridge CB2 0QH UK email: jyl(a)mrc-lmb.cam.ac.uk website: http://www2.mrc-lmb.cam.ac.uk/groups/jyl

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Re: [phenixbb] rotamer outlier and clash socre
by Pavel Afonine 14 Feb '21

14 Feb '21

Hello Pan, """ I refined a cryo-EM data with 3.2 angstrom, the model fit well with the map, but the rotamer outlier is too high, about 8%. How can I decrease the value of rotamer outlier by realspace.refinement in Phenix GUI, and no need to adjust the residues one by one in Coot? """ if you used latest version of Phenix, which is dev-4146 as of today: http://phenix-online.org/download/nightly_builds.cgi?show_all=1 then this is not expected, because phenix.real_space_refine has built-in ability to detect which residues need a fix and does an exhaustive grid-search in chi-torsion angle space to optimize it against the map and local environment. If this happened using the latest version of Phenix, then this is an indication to me that something has gone wrong and we need to investigate. For this I'm asking to send all input files. Residues that phenix.real_space_refine was unable to fix for whatever reason are best to fit manually in Coot. And I can't agree more with Tristan -- all the great tools and automation are to help you not substitute you! So, checking manually is important! """ Another question is the value of clash score are also too high. """ You did not tell us the value, so no answer here! A general guidance: the lower the better, but zero is not the goal. """ When I set the nonbonded_weight to 10000 """ According to study we conducted recently: http://phenix-online.org/presentations/nb_weight.pdf this unlikely to be a great idea. Pavel

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rotamer outlier and clash socre
by xiaowp82＠hotmail.com 14 Feb '21

14 Feb '21

Hello everyone I refined a cryo-EM data with 3.2 angstrom, the model fit well with the map, but the rotamer outlier is too high, about 8%. How can I decrease the value of rotamer outlier by realspace.refinement in Phenix GUI, and no need to adjust the residues one by one in Coot? Another question is the value of clash score are also too high. When I set the nonbonded_weight to 10000, the clash score get better, but the value of rotamer outlier get worse. The high clash score maybe due to including hydrogens for refinement? Can I exclude hydrogen for refinement and how to do? Thanks ! Pan xiaowp82(a)hotmail.com

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Phenix ligand identification resolve was not able to finish on the first cycle
by Ahmad Khalifa 14 Feb '21

14 Feb '21

Hello, I want to use phenix.ligand_identification to identify a ligand. I used chimera's volume eraser to erase everything except for the ligand density. I then converted it to mtz. in the phenix.ligand_identification gui, I set reflection data type to difference map. When I ran the job, I got a lot of "Sorry, resolve was not able to finish on the first cycle". Regarding the mtz, I have to contour the map to 170rmsd to visualize it in coot, so something have to be wrong. I appreciate any input. Best regards.

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