Dear Jeremy, Indeed, we also incline to think of it as a monomer in solution, but still quite un-eased by the extensive interactions in the asu being merely as a result of a crystallization artifact. As you said, we may need to rely more heavily on biochemical analysis and since SEC wasn't clear we are turning now to LS (hope to able to post a more conclusive update).
Regardless of what our final conclusion would be for this case, we became rather generally interested to find other similar cases of homomeric assemblies related only by non crystallographic translation symmetry (or as Engin Qzka pointed out "improper NCS" is the conventional terminology). So to rephrase our question we are interested to learn about additional structures of homomoeric improper ncs assemblies. I truly appreciate ANY open-minded or skeptic thought, profound or trivial that we get here! They all, definitely those made by Mark Garavito, contribute to shaping our mind around this riddle. Thanks for commenting on the skepticism, I brought it up as part of the discussion but a glitch of my own coffee time haziness might have slipped in. Perhaps I should try some o-cha instead .. :) cheers, Hay On Dec 12, 2014, at 3:05 AM, Jeremy Tame wrote: > Dear Hay > > I suggest that you use analytical ultracentrifugation to determine the > oligomeric state of the protein in solution. > Mass spectrometry and light scattering are also useful, but there are so many > examples of gel filtration proving > erroneous it has questionable value as an analytical technique. For an > example of a dimer interface predicted by > PISA to be real you could look at Yoshida et al, JMB 423, 351 (2012). The > protein is in fact a monomer in solution. > PISA is a fantastic tool, but interfaces in crystals do not always reflect > the solution state. My guess (with the > information I have) is that your protein is probably a monomer too. > > With regard to Michael Garavito's reply requesting more information, I would > like to comment that scepticism > is indeed an important god in the pantheon of science, but that that minor > deity open-mindedness also deserves the > occasional nod. 10-fold crystal symmetry is one example, but the list of > "impossible" things now become mainstream > is a long one (continental drift, Earth >100,000 years old, quantum > mechanics....and so on). Bayes theorem cannot > help you discover the truth if you have set its prior probability to zero. > But I haven't my morning o-cha yet either. > > good luck > Jeremy > > > On Dec 11, 2014, at 9:27 PM, Hay Dvir wrote: > >> Dear all, >> >> >> We have a structure of a rather tightly packed homotrimer protein in the ASU >> with no apparent crystallographic or non-crystallographic rotational >> symmetry between monomers. >> Attempting to establish the biological assembly, we are very interested to >> hear about additional similar cases you might know of. >> >> Thanks in advance, >> Hay >> >> >> --------------------------- >> Hay Dvir Ph. D. >> Head Technion Center for Structural Biology >> Technion Haifa 3200003, Israel >> Tel: +(972)-77-887-1901 >> Fax: +(972)-77-887-1935 >> E-mail hd...@technion.ac.il >> Website http://tcsb.technion.ac.il >> >
