I have done once by using two proteins, one is disordered, the other is very well folded. The result I got is the baseline drift. The baseline goes up upon each injection. The reason I thought at that time is the heat capacity changed dramatically in the system. The disordered protein may form some degree of structure after interacting with its partner. And we did find some precipitation after the experiment.
Dee Xiaodi.yu@childrens.harvard edu Sent from my iPad > On Mar 14, 2014, at 9:57 AM, "Anita P" <crystals...@gmail.com> wrote: > > 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> 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 >> >> >> ---- Original message ---- >> >Date: Fri, 14 Mar 2014 18:07:48 +0530 >> >From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> (on behalf >> of Anita P <crystals...@gmail.com>) >> >Subject: [ccp4bb] ITC with unfolded proteins >> >To: 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 >
