Hi Peter
This idea was discussed at the recent RAMC meeting, and there is at least
one example where it has worked.
Generally, cross-seeding can work as long as you have homology. See e.g.
Obmolova et al. Acta Crystallogr. 2010, D66, 927–933. The same group has
reported seeding a complex with
agree, any crystallisation idea is worth pursuing, given you have or can make
enough sample to try it with.
having said that, wouldn't you tend to select for the same crystals as the
seed, i.e. crystals of the component on its own?
have you tried limited proteolysis of your sample, incl. a bit
On Wed, 2011-09-21 at 18:04 +0100, Peter Hsu wrote:
Or is this just a crazy/bad idea?
If there is one thing that I learned about crystallization, is that very
few ideas are so crazy that they are bad (i.e. not worth trying). Well,
if dried seaweed and ground horse hair are good for seeding, I
I forgot to mention, I can reconstitute the complex (co-expression/mixing
proteins) and the complex comes off both ion exchange and an SD200 as one peak.
Just haven't had luck with getting crystals.
The idea is not at all crazy. In a sense it is quite similar to
Stoichiometric variation screening* if you consider that the lattice of
the crystallized subunit may contain planes
that might be conserved in the crystal of your hope for 3 protein complex.
*Stura, E.A., Graille, M., Taussig,
agree, any crystallisation idea is worth pursuing, given you have or can make
enough sample to try it with.
having said that, wouldn't you tend to select for the same crystals as the
seed, i.e. crystals of the component on its own?
have you tried limited proteolysis of your sample, incl. a
