Hello,
If you Google Alan cooper ITC insulin, you should find his work describing the study of insulin dimer:monomer equilibrium and the effect of cyclodextrin studied via ITC. This may be of some help.
Cheers
Mike
From: CCP4 bulletin board <[log in to unmask]>
On Behalf Of Bernhard Rupp
Sent: 03 October 2019 16:52
To: [log in to unmask]
Subject: Re: [ccp4bb] ITC question -dimer vs monomer
I am not looking for anything yet – I wonder what – if any – the consequences of doing it one way or the other would be.
I am reasonably certain that any difference affects the analysis.
Thx, BR
From: CCP4 bulletin board <[log in to unmask]>
On Behalf Of Keller, Jacob
Sent: Thursday, October 3, 2019 17:41
To: [log in to unmask]
Subject: Re: [ccp4bb] ITC question -dimer vs monomer
I don’t understand what you are trying to do—are you trying to show, by the difference in ITC response, that the predictions you made about the oligomerization are true?
JPK
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From: CCP4 bulletin board <[log in to unmask]>
On Behalf Of Bernhard Rupp
Sent: Thursday, October 3, 2019 11:06 AM
To: [log in to unmask]
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
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Bernhard Rupp
http://www.hofkristallamt.org/
+1 925 209 7429
+43 676 571 0536
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Many plausible ideas vanish
at the presence of thought
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