Do you have an inactive variant of the protein? If yes, use limited
proteolysis in the presence of methylated histones to find out which
flexible parts are really important and or protected by complex
formation. Further, try to crystallize a complex of inactive demethylase
and methylated histones, a long shot but a good way to combat the
apparent flexibility....
Good luck,
Jeroen
Am 04.07.17 um 17:22 schrieb dongxiaofei:
Dear ALL,
I want to make a protein crystal,but there is a long loop between
domains of protein , which contains two small domains owning about 40
amino acids respectively and a loop about 70 amino acids.
Loop is so long and flexible ,but I don't want to delete some
fragments,because it may be important for protein's function of a
histone demethylase.
Besides, the surface charge of protein is whole negative .
I have tried a long time but it is hard to me to get crystal.
Would be very grateful for any advice!
Thanks
Dong Xiao
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Dr.math. et dis. nat. Jeroen R. Mesters
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It is invariably the case that high resolution X-ray structures show
significantly better agreement with solution observables such as
coupling constants, 13C chemical shifts, and proton chemical shifts,
than the corresponding NMR structures, including the very best ones.
Hence, in most cases, a high-resolution crystal structure (< 2.0 Å)will
provide a better description of the structure in solution than the
corresponding NMR structure (Kuszewski, Gronenborn & Clore, 1996,
Protein Science 5:1067-80)
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