[ccp4bb] Spam : Re: [ccp4bb] Off-topic: Attach His-tagged Protein to Coverslip

Assuming the protein is in solution, drop cast 2ul of solution onto a glass surface and spread evenly with a sharp needle. Air dry in dust free chamber and blow the surface with gentle stream of nitrogen gas. Proteins stick to surface quite well to be analyzed for various microscopy (AFM, SEM

[ccp4bb] Off-topic: Attach His-tagged Protein to Coverslip

Does anyone have a simple way to attach purified his-tagged protein solidly to a coverslip? Thanks, Jacob Keller *** Jacob Pearson Keller, PhD Research Scientist HHMI Janelia Research Campus / Looger lab Phone: (571)209-4000 x3159 Email:

[ccp4bb] Postdoctoral position in Sydney, Australia

Dear CCP4bb members, We are seeking a recent PhD graduate for a postdoctoral position in structural biology. Applicants with experience in protein crystallography as well as mammalian cell culture, computational biology or RNA biology are particularly encouraged to apply. Please see link below

[ccp4bb] Accidental crystallization of E. coli protein

During the crystallization of a totally unrelated protein from a different bacterium in E. coli, we managed to somehow crystallize an E. coli protein. It turned out to be only the catalytic domain of an enzyme. Two previous reports both used recombinant expression of this enzyme followed by

Re: [ccp4bb] E.coli strain, knock-out for SDH/FR

People who would know (and know whether such a strain would even be viable) are Gary Cecchini at UC San Francisco/VA and Bob Gennis at Univ of Illinois at Urbana. In case you are not already talking with them. It has been reported that an assembly factor (SDH5 or SDHE or SDHAF2) is required for

Re: [ccp4bb] Helium-Temp Cryo-Cooling

Hi Jakob, There are several other examples where the effect of temperature on radiation damage for metallo proteins was demonstrated, eg: 10.1074/jbc.M112.438796 10.1073/pnas.0505207102 10.1074/jbc.M509724200 The tricky part with liq He cryo streams is to get a good geometry that provides stable

Re: [ccp4bb] Using a codon-optimised gene to improve proteinsolubility

Dear Sutapa, As already pointed out, codon optimization for increasing the solubility is not really a viable option but it can be for production /per se/ in insect cells. Among others, slowing down growth, changing the construct or coexpression (of for example foldase/chaperone) or witching

[ccp4bb] anomalous difference fourier

I suppose I should have expected this, but I still was led astray.. Ir atom - strong anomalous signal.. Good peak in anom diff fourier phased from protein. So I add it to the model with occ = 1.00 and refine.. The B factor is very high and it seems clear that the occupancy should be <<1.0 And

Re: [ccp4bb] Using a codon-optimised gene to improve protein solubility

Hello Sutapa, With codon optimisation you would get more and faster expression and likely even more insolubility in E. coli (less time for folding). To get the protein more soluble perhaps slower production might help. I guess you could try codon de-optimisation :-). Cheaper possibilities are

Re: [ccp4bb] Ambiguous crystal diffraction pattern

It’s multiple crystals of salt. JPK From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Prashant Deshmukh Sent: Tuesday, April 04, 2017 8:40 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Ambiguous crystal diffraction pattern Dear Crystallographers, Please help us in figuring

[ccp4bb] rapper

Dear all, Has anyone tested RAPPER recently - doenst seem to work with me from the ccp4i 7.0.015 with a map (from buster-TNT). (either ca-trace option - with partial region defined or the "rebuild badly fitting residues" - complains and generates nothing. Is there something obvious I might be

Re: [ccp4bb] Helium-Temp Cryo-Cooling

Jacob Good question. One case I know is for a metalloprotein where the photoreduction of the metal centre is decreased significantly at 40K compared with 110K. Acta Cryst. (2007). D63, 951-960