Dear Anita One alternative method to determine the thermodynamics and potentially discern the folding energy changes from the interaction driven ones would be NMR. Advantages over ITC experiments include: determine if the interaction drives foldness, estimate associated thermodynamics and, if you do 3D labelling experiments and do backbone assignment, you can map the interaction interface from the NMR data. Elegant NMR experiments have been extensively exploited to study unfolded/folded states and interactions in recent times, with exciting results which proved to be unachievable by any other biochemical/biophysical/structural technique, so it was definitely worth trying!
Good luck Paula =================================== Dr Paula S. Salgado Lecturer in Macromolecular Crystallography Institute for Cell and Molecular Biosciences Faculty of Medical Sciences 3rd Floor Cookson Building Newcastle University Newcastle upon Tyne, NE2 4HH, UK Tel: +44 (0)191 222 7369 Fax: +44 (0)191 222 7424 Email: paula.salg...@ncl.ac.uk ________________________________________ From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Anita P [crystals...@gmail.com] Sent: 14 March 2014 13:57 To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] ITC with unfolded proteins That is a very interesting question, which I would request the seniors out there to give their insights on. I was imagining that a recombinant purification of an unfolded partner would aggregate which would cause trouble in ITC. Am I correct in this theory? Would love to have more insights. thanks in advance Anita On Fri, Mar 14, 2014 at 7:18 PM, <rkha...@ccny.cuny.edu<mailto:rkha...@ccny.cuny.edu>> wrote: Hi, I think the experiment is doable, but how would you decouple protein-protein interaction from folding of the unfolded protein due to protein interaction? Reza Reza Khayat, PhD Assistant Professor The City College of New York Department of Chemistry, MR-1135 160 Convent Avenue New York, NY 10031 Tel. (212) 650-6070<tel:%28212%29%20650-6070> ---- Original message ---- >Date: Fri, 14 Mar 2014 18:07:48 +0530 >From: CCP4 bulletin board ><CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK>> (on behalf of Anita P <crystals...@gmail.com<mailto:crystals...@gmail.com>>) >Subject: [ccp4bb] ITC with unfolded proteins >To: CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK> > > Hello everyone, > I have a query for the scientists working on > protein-protein interaction. > It is known that some proteins exist in unfolded or > molten globule state and attain structure on > interaction with other folded proteins. > Many a times, it is difficult to obtain the > structure of these complexes. > Is it possible to quantitatively determine the > thermodynamics of interaction between an unfolded > protein and a folded protein using ITC? Later may be > perform an alascan to determine the residues of the > unfolded partner involved in the interaction. > Please share your ideas > cheers** > Anita
