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 <[email protected]> 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: [email protected] *******************************************
