Dear Dr.BR There could be two possibilities, you could try to do titration of dimer into monomer loaded in cell or vice versa, and with subtraction of heat of dilution from protein-protein titration. These kind of molecular interaction involving distinct oligomer units need be addressed by any other orthogonal methods.
Regards Rajivgandhi Sundaram On Thu, Oct 3, 2019, 10:21 PM clare stevenson (JIC) < clare.steven...@jic.ac.uk> wrote: > I would try it both ways and see what you get. Also do controls of buffer > into each protein > > > > For extra info could also try with SPR. Always best to do these things > using multiple complimentary methods > > > > Best wishes > > > > Clare > > > > *From:* CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> *On Behalf Of *Bernhard > Rupp > *Sent:* 03 October 2019 16:06 > *To:* CCP4BB@JISCMAIL.AC.UK > *Subject:* [ccp4bb] ITC question -dimer vs monomer > > > > Hi Fellows, > > > > please let me ask the respective experts an ITC question: I have 2 > proteins, stable and dialyzed in identical buffer. > > A is a monomer and B an obligate dimer. I suspect that eventually a A2B2 > dimer will form. > > > > Intuitively, it should make a difference whether I titrate the dimer with > the monomer or vice versa. > > In the first case, a momomer would initially meet a lot of free dimers, > and I would expect that randomly, a AB2 complex > > is more likely to form than a A2B2 (let’s disregard any more complex > colligative/cooperative effects). > > > > If I drip the dimer into the monomer pool, it is quite likely that the B > dimer meets 2 free As, and I get right away a higher population of A2B2s. > > > > Maybe at dilutions of ITC and with sufficient equilibration that is not an > issue at all (again, absent any cooperative effects that might alter the > first Kd vs. the second, despite the sites on the dimer are at least > initially equivalent). > > > > Can someone guide me towards literature about this or perhaps share some > first-hand experience? > > > > Many thanks, BR > > > > ------------------------------------------------------ > > Bernhard Rupp > > http://www.hofkristallamt.org/ > > b...@hofkristallamt.org > > +1 925 209 7429 > > +43 676 571 0536 > > ------------------------------------------------------ > > Many plausible ideas vanish > > at the presence of thought > > ------------------------------------------------------ > > > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
