[ccp4bb] Senior Scientist, Structural Biology position at Sanofi US - Boston area
We are seeking a Scientist to join our Boston-area structural biology group conducting drug design research in diverse therapeutic areas. Our group integrates with multidisciplinary project teams to accelerate drug discovery on innovative and challenging targets using x-ray crystallography and cryo-EM. To expand the latter, a state-of-the-art facility is under construction at our new 2022 Cambridge campus. The successful candidate will generate structure-grade protein, and use such protein for structural studies using cryo-EM or crystallography in order to drive drug-design workflows. Significant experience with single-particle cryo-EM data generation and processing is desired. The successful candidate will work closely with members of the Structural Biology, Biochemistry and Biophysics teams, as well as members of the therapeutic area groups, and must be an enthusiastic team player with excellent communication skills and must bring curiosity and a desire to make an impact in the workplace. Basic Qualifications: PhD in Biochemistry or a related field (Biology, Cell Biology, Biotechnology, etc.) and 3+ years of experience in protein expression, purification and cryo-EM studies, or BS degree and 8+ years of relevant experience. Preferred Qualifications: Extensive track record with purification and structure determination of challenging proteins, (e.g., membrane proteins, complexes, etc.) by cryo-EM or crystallography. Hands-on operation of high-end cryo-electron microscopes, extensive familiarity with EM data processing suites. Industrial experience. If interested, please apply using this link: https://sanofi.wd3.myworkdayjobs.com/SanofiCareers/job/Waltham-MA/Senior-Scientist---Structural-Biology_R2621212-1 -- Michael Kothe Senior Principal Scientist Structural Biology, Sanofi R&D 153 Second Avenue Waltham MA 02451 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
[ccp4bb] Emerald Plugmaker experiences?
Hi, I am looking for feedback from those who have used the Emerald Plugmaker instrument, especially if in your experience the translation of initial crystallization hits to diffracting crystals was as straightforward as claimed by Emerald. Any other thoughts and comments are also welcome. Thanks, Michael
Re: [ccp4bb] protein stain, B-PER
Hi Artem, I have stayed away from these types of reagents in order to avoid detergents, and also because of anecdotal reports that these work less well for large-scale preps. Are you saying you use them routinely for preps of crystal-grade protein? Am I being too concerned about the presence of detergents? Thanks, Michael From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Artem Evdokimov Sent: Thursday, March 15, 2012 7:04 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] protein stain, B-PER Hi :-) These subjects come up regularly! Basically, in brief we use a nearly magical mix of bper and yper in 3 to 1 ratio, supplemented with salt, bensonaze and lysozyme. In my fairly extensive experience this works for about 85 to 90 % of cytoplasmic proteins with no issues for affnity chromatography and (if no salt is added) for ion exchange primary capture. For the remaining 10% we try every other method under the sun. For gels - 15 minute biorad gels are very nice (have to have 300v power supply) and stain/destain in our hands is abou 5-15 minutes with a microwave. We destain in boiling water, with no other chemicals involved. Any coomassie based stin works well enough for this. Some ppl in our lab love the stain free gels but you have to remembr to count trp residues as it relies on trp chemistry to work. In short, this (and a number of other tricks) makes it possible to have a three step purification done in one 8 hr work day. Artem On Mar 15, 2012 10:25 AM, "Thomas Edwards" wrote: Dear BB, Apologies for being mildly off topic. Maybe. 1. We are trying to express (in E. coli) a protein which appears to be quite sensitive to mechanical disruption. We have ordered some B-PER (Pierce - "B-PER Bacterial Protein Extraction Reagents are designed to extract soluble protein from bacterial cells without harsh chemicals or mechanical procedures like sonication"), but would like to try a variety of similar things if possible. Any advice from the community out there? Anybody know what goes into B-PER or similar things (I know there's some Dnase and lysosyme in there - but which detergents are compatible with Ni, GST, how much do you need etc)?? 2. Staining SDS gels. There are various concerns from lab members about safety re methanol in stains, microwaving stains etc etc. "Instant Blue" claims to have none of these problems. Quote: "Protein gel staining takes around 15 minutes without the need to wash, fix, microwave or destain". But again, I'd like to try things to see if they work for us (before spending cash - yes, I am spending averse...!). Anybody any suggestions for quick, non-fix, non-methanol, non-microwave, non-destain protein gel stains? Have tried home made colloidal coomassie but our protocol still requires fixes and washes that made it not really worth while. Happy to collate thoughts on replies offline and post summary. Many thanks Ed T.A.Edwards Ph.D. Deputy Director Astbury Centre for Structural Molecular Biology Lecturer in Biochemistry Garstang 8.53d University of Leeds, Leeds, LS2 9JT Telephone: 0113 343 3031 http://www.bmb.leeds.ac.uk/staff/tae/ -- No one should approach the temple of science with the soul of a money changer. ~Thomas Browne
Re: [ccp4bb] Using chaperones to boost expression in E. coli
Hi, I wanted to thank everyone for their responses to my inquiry. It's good to see that there have been positive examples, and also that one should not be discouraged from trying by initial negative results. Thanks again, and have a nice weekend, Michael From: Olga Kolaj [mailto:olga.ko...@gmail.com] Sent: Tuesday, April 26, 2011 5:17 AM To: Kothe, Michael Cc: CCP4BB@jiscmail.ac.uk Subject: Re: [ccp4bb] Using chaperones to boost expression in E. coli Michael, We have put together a review about use of folding modulators to improve expression in E. coli some time ago which might be useful to you (Kolaj et al Microb Cell Fact. 2009 Jan 27;8:9). Here is the link: http://www.ncbi.nlm.nih.gov/pubmed/19173718 <http://www.ncbi.nlm.nih.gov/pubmed/19173718> Regards On Fri, Apr 22, 2011 at 4:28 PM, Kothe, Michael wrote: Dear ccp4bb, I am curious to hear of examples where the expression of well-behaved protein was achieved by the coexpression of chaperones in E. coli. I know the appropriate strains and vectors exist, but I can't remember hearing of a successful case. I have heard anecdotally of several cases where it was tried without success (including one attempt I made myself). I also heard of concerns that the chaperones might copurify with the (now soluble) protein of interest and are difficult to remove. Thanks, Michael
[ccp4bb] Using chaperones to boost expression in E. coli
Dear ccp4bb, I am curious to hear of examples where the expression of well-behaved protein was achieved by the coexpression of chaperones in E. coli. I know the appropriate strains and vectors exist, but I can't remember hearing of a successful case. I have heard anecdotally of several cases where it was tried without success (including one attempt I made myself). I also heard of concerns that the chaperones might copurify with the (now soluble) protein of interest and are difficult to remove. Thanks, Michael
[ccp4bb] Postdoctoral opportunity in Structural Biology at Genzyme Corporation in Waltham MA
Genzyme Corporation, ranked as one of the foremost biotechnology companies in the world, is committed to providing an exceptional environment in which individuals can excel, and achieve their professional and personal goals. Genzyme Corporation has been selected by FORTUNE magazine as one of the "100 Best Companies to Work For in 2006 in the United States". By applying for a position with Genzyme, you are taking the first step toward becoming a part of our dynamic and talented team, and hopefully sharing in our continued success. We have a postdoctoral position open in the Fragment-Based Drug Discovery (FBDD) group in Drug and Biomaterial R&D at Genzyme Corporation in Waltham, MA. We are seeking a highly motivated recent PhD graduate in protein X-ray crystallography for a two-year position with a possible one-year extension. This position offers opportunities to work on structural biology of protein targets of therapeutic interest for FBDD approach, which involves activities from construct design to structural determination of target protein in complex with substrates or inhibitors. The successful candidate will have access to our cutting edge FBDD technology, including state of the art crystallization equipment and X-ray diffraction system. Basic Qualifications: PhD in structural biology or related disciplines (biochemistry, biophysics) with track record of solving X-ray crystal structures of proteins and protein complexes. Experience with construct design, protein purification, protein crystallization, data collection, structure determination and interpretation are essential. Preferred Qualifications: Strong background in biochemistry with in-depth understanding of ligand-protein interactions and familiarity with techniques such as ITC, surface plasmon resonance etc. to characterize proteins in support of structural biology is highly desired. Please submit your application on l...@www.genzyme.com
