Hey guys I'm working with an extremely flexible multi-domain protein which has proved highly resistant to crystallization. I've tried a variety of concentrations, protein-ligand concentrations and screens with no success. Is there a particular set of conditions that is more conducive to crystallising flexible proteins?
Best regards
Harry
Hi Harry,
Of cours, you can try different buffers conditions.
But, in order to optimize your chance to get a crystal, you will have to regidify your protein:
- either by finding a high affinity ligand able to maintain the structure,
- or by mutating the flexible region (usually by reducing the length). Of course, this way has to be justified in regard of the protein function.
Good luck for your project,
Yves
Hi Harry,
a new kit for determination of stability of protein (- and crystallization conditions) is the Durham pH Screen which is now available. This might also help with flexible proteins.
Hth and good luck,
Stefanie
Hi there,
The key would be to limit the flexibility of the whole molecule. In case natural partners are not known, on approach is to generate artificial protein partners by directed evolution, form the complex and solve the structure of the complex.
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