I probably it depends on whether you've got gunk or a functionally relevant oligomer in that void volume. Is it active?
RecA and Rad51 both form open-ended head-to-tail oligomers and yet they still crystallize. ===================================== Phoebe A. Rice Dept. of Biochemistry & Molecular Biology The University of Chicago phone 773 834 1723 http://bmb.bsd.uchicago.edu/Faculty_and_Research/01_Faculty/01_Faculty_Alphabetically.php?faculty_id=123 http://www.rsc.org/shop/books/2008/9780854042722.asp ---- Original message ---- >Date: Tue, 21 Feb 2012 18:21:03 -0500 >From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> (on behalf of Raji >Edayathumangalam <r...@brandeis.edu>) >Subject: [ccp4bb] Aggregated protein for crystallization >To: CCP4BB@JISCMAIL.AC.UK > > 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
