Hey, it could be that you just have a big oligomer--any support for
that in the relevant literature? A 10-mer would probably beat out an
s200, no? Do you have any other way to ascertain the oligomeric state?
Jacob
On Tue, Feb 21, 2012 at 5:21 PM, Raji Edayathumangalam
<[log in to unmask]> wrote:
> Hi Folks,
>
> As crazy as it sounds, if you have crystallized and managed to solve the
> structure of a protein from aggregated protein, please could you share your
> experience.
>
> After many constructs, many many expression schemes and after the usual
> rigmarole of optimization that is also often discussed on ccp4bb (buffers,
> glycerol, salt concentrations, pH, detergent, additives etc.), I now have a
> decently expressing truncated construct for my protein (80 kDa) that is pure
> but aggregated (elutes in the void volume from a Superdex200 column). I am
> tempted to make a boatload of aggregated protein and set up some crystal
> trays (after perhaps testing by CD). So I'd like to hear from folks who have
> been successful in solving structures from aggregates when many many known
> and tested optimization methods still leave one with aggregated protein.
>
> Thanks.
> Raji
>
> --
> Raji Edayathumangalam
> Instructor in Neurology, Harvard Medical School
> Research Associate, Brigham and Women's Hospital
> Visiting Research Scholar, Brandeis University
>
>
--
*******************************************
Jacob Pearson Keller
Northwestern University
Medical Scientist Training Program
email: [log in to unmask]
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