[ccp4bb] tool for calculating average alpha helical rise per trun
Hello could any suggest some ways/tools to calculate average alpha helical rise per turn for a helical model from PDB. Thank you in advance regards uday
[ccp4bb] CCP4 Newsletter 48
Hi folks, After a brief hiatus (okay, more than four and a half years) the CCP4 Newsletter on Protein Crystallography is back! The latest issue is now available from the website http://www.ccp4.ac.uk/newsletters.php in both HTML and PDF. This edition follows hot on the heels of the 6.3.0 release of the CCP4 Suite, and correspondingly it focuses mainly on new and improved programs found in the Suite. There are articles on the new programs ViewHKL, GESAMT, comit, nautilus, ProSMART, Zanuda and AMPLE. We also have news of the latest improvements to the mosflm package and the current xia2 manual. Finally, two articles about the new GUI project, CCP4i2, give a taste of things to come from CCP4 developers. We intend to get back to a semi-regular schedule of Newsletter publications. To that end, submissions are accepted immediately for the next issue, with a target publication date in the winter. Articles may cover any topic of interest to macromolecular crystallographers, though we are particularly interested in articles on software or methodology, and short items of news. Have a good weekend. -- David Waterman
[ccp4bb] best strategy to establish stable expression in mammalian cells
Dear All; We are planning to establish stable CHO or 293 cell clones to express some receptors. I heard that it is time consuming to select high expression clones using pcDNA3.1 vectors. As our crystallographer are dealing with all kinds of proteins, can anyone recommend a best strategy for mammalian cell expression? Thank you so much and have a nice weekend, Jerry McCully
Re: [ccp4bb] best strategy to establish stable expression in mammalian cells
Hi Jerry, Perhaps this Nature protocols paper from the Stroud Lab at UCSF might be useful. Overexpressing human membrane proteins in stably transfected and clonal human embryonic kidney 293S cells http://www.nature.com/nprot/journal/v7/n3/full/nprot.2011.453.html -john On Aug 17, 2012, at 3:38 PM, Jerry McCully wrote: Dear All; We are planning to establish stable CHO or 293 cell clones to express some receptors. I heard that it is time consuming to select high expression clones using pcDNA3.1 vectors. As our crystallographer are dealing with all kinds of proteins, can anyone recommend a best strategy for mammalian cell expression? Thank you so much and have a nice weekend, Jerry McCully
Re: [ccp4bb] Evaluating crystallogarphic experiment
Quite common. And it may be a good sign for crystallizability in those conditions. As the concentration gradients dissipate from the initially mixed drop, your protein re-solubilizes. Roger Rowlett On Aug 17, 2012 5:31 PM, Yuri Pompeu yuri.pom...@ufl.edu wrote: Hello everyone, I have just made a curious observation. I purified an enzyme domain and concentrated it to around 16 mg/mL without much mischief. I then proceeded to set-up some drops. After 5 min most of the drops had a muddy appearance to them which led me to think I was probably too concentrated in either protein and/or precipitant. Curiously I went back to look at the drops after an additional 30 min and they all look pretty clear with no appreciable precipitation. Has anyone encountered this situation or a similar one before? Any input/shared experience is welcome. Best, Yuri
