Dear Martin, A more bleeding-edge type of experiment in the vein David is indicating would be to use a "rotation SFX" protocol to record complete data off one or very few crystals that will be completely exempt from radiation damage, as described in doi:10.1038/nmeth.2962 and in doi: 10.1073/pnas.1418733111 .
Good luck! Gerard -- On Mon, Nov 06, 2017 at 08:07:40AM -0500, David Schuller wrote: > Some alternative interpretations have been suggested, but if you think you > are seeing radiation damage, you could try collecting data on several > crystals and binning it by dose received. For comparison see: > > > The catalytic pathway of horseradish peroxidase at high resolution. > Berglund, et al. (23 May, 2002) Nature 417(6887), 463-8 > DOI: > 10.1038/417463a <https://doi.org/10.1038/417463a> > > > If your low dose and high dose structures appear nearly the same, then you > are seeing something other than radiation damage. > > > > On 11/06/17 06:01, Martin Malý wrote: > > Dear colleagues, > > > > I am investigating a structure of a FAD-dependent enzyme. The electron > > density map suggests radiation damage to the FAD. It apparently is > > different from simple change of the redox state and "butterfly"-like > > structure. We did not find in literature possible products of radiation > > damage, like a removal of several atoms of the FAD. Has anyone observed > > such effect? > > > > To describe it in more detail, I can observe negative difference map of > > C2, N3, C4, and O4 atoms of flavine. Moreover, there is positive > > difference map close to O2 and O4 atoms thus it looks as water molecules > > are bound there instead of the missing FAD atoms. > > > > I am attaching parameters of the experiment: performed at synchrotron, > > exposition time 210 s, high resolution diffraction limit 1.65 A > > (<I/sigma> = 2 at shell 1.75-1.65 A). We could see a decrease of > > diffraction data statistics during the experiment hence we think there > > is significant radiation damage to the crystal. > > > > Thank you very much for ideas. > > Regards, > > Martin Maly > > > -- > ======================================================================= > All Things Serve the Beam > ======================================================================= > David J. Schuller > modern man in a post-modern world > MacCHESS, Cornell University > schul...@cornell.edu >