Spherulites....

Artem Evdokimov AEVDOKIMOZ at cinci.rr.com
Fri Oct 5 21:14:59 EST 2001


What's the protein like ? Size ? Is it post-transcriptionally modified ? Does it look like it has floppy bits ? What's the pI ? 

Questions are endless, and so can be the suggestions :) Obvious things to try include: changing temperature.  Removing modifications. Adding modifications. Limited proteolysis. Heavy atom salts in screens. Mutagenesis a-la Derewenda et al. Using NDSB's and/or detergents.

Good luck !

A.
  "Mike Hanlon" <michael.hanlon at bbsrc.ac.uk> wrote in message news:3BBDC7C5.EDB2EA65 at bbsrc.ac.uk...
  Spherulites can be a good sign - so try setting up an optimisation screen. Varying the PEG concentration by a few % and the pH by 0.1steps around the original conditions. Even a slight change of conditions can make the difference between xtal and no xtal 
  Good luck 

  Debashis Mukhopakhyay wrote: 

      
Hi all,
I'm looking for some suggestions regarding the following problem.
After prolonged trial I got some spherulites for a protein I'm
trying to crystallize, the conditions being 25-30% PEG300 in Citrate
(pH 5.5). They are solid (crystal-like) to touch and also takes the
Izit-dye (Hampton). I'm looking for ideas as to what to do next to
get a nice diffracting crystal.
Sharing of any experience in the matter would be highly appreciated.
Thanks,
Deb 
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