One of my main concerns is that the unfolded protein itself would irreversibly aggregate and then wouldn't interact with the folded protein. I think DSC (differential scanning calorimetry) should be performed first to characterize the state of both proteins and their potential "complex". By analyzing the DSC data, you should have an idea what kind of enthalpy change is involved during the complex formation. If you see the unfolded protein does become folded, then ITC experiments are worth trying. A combination of ITC/DSC experiments may help you tweak out the exact interaction parameters.
Wendy -------------------------------------------------- Wendy Yang Manager, Biocalorimetry Laboratory Center for Biophysical Sciences & Engineering University of Alabama at Birmingham Tel: 205-975-2450 Fax: 205-934-0480 Email: y...@cbse.uab.edu<mailto:y...@cbse.uab.edu> ________________________________ From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Anita P Sent: Friday, March 14, 2014 8:57 AM 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
