If the following is not deleterious to your protein and its function you could introduce mutations that prevent dimerization.
~Jeff > I have a similar case, where in there are multiple binding sites on the > protein for the ligand and ligand induces dimerization.So it is not > helpful > even if I separate the monomer and dimer. > If I titrate the dimer with ligand, the stoichiometry will completely > change? Any suggestions will be helpful. > > > On Fri, Jul 18, 2014 at 12:46 PM, David Briggs <drdavidcbri...@gmail.com> > wrote: > >> Hi Sajid, >> >> *Assuming* you have one site per monomer (rather than, say, one site per >> dimer), and *assuming* each binding event is completely independent ( >> I.e >> no co-operativity), you might just get away with running the experiment >> with the heterogeneous material. >> >> However, you might not be able to confidently make these assumptions, so >> imho it would be preferable to separate the monomer and dimer by SEC >> prior >> to ITC. If this is not possible, then pay close attention to the fit >> when >> you run the heterogeneous experiment. Poor fit to a one site model may >> indicate that these assumptions are invalid. Can you obtain >> stoichiometry >> information from a different technique? This might be very helpful. >> >> Hth, >> >> Dave >> >> Dr David C Briggs PhD >> http://about.me/david_briggs >> On 18 Jul 2014 10:25, "sajid akthar" <b_sajid_...@yahoo.co.in> wrote: >> >>> Dear All, >>> >>> This is an off-topic question. I have protein solution of heterogeneous >>> (contains both monomer and dimer). I want to perform ITC with this >>> protein. >>> I doubt whether this heterogeneity will interfere the binding study. >>> >>> Any advice please. >>> >>> Thank you >>> >>> Sajid >>> >> >
