[Myotox] myotoxin2-SDS titration

[Borries Demeler]

AUC is not a choice method for studying kinetics since the time scale of
sedimentation is too slow, except for very large molecules and high rotor
speeds. Myotoxin is only 13.5 kDa or so, therefore there isn't any kinetics
you could pick up. But you can measure Kds very well, which is what we are
doing now. I'll explain more on Tuesday.
-b.

On Thu, Apr 1, 2021 at 6:16 PM Hazendonk, Paul <paul.hazendonk at uleth.ca>
wrote:

> So if you have association-dissociation taking place, the slope at the
> inflection point does not change with speed.  I guess the exchange rates
> are too high.  How about for very slow processes?  I guess at that point
> you would see separate inflections.
>
> -----Original Message-----
> From: Myotox <myotox-bounces at biophysics.uleth.ca> On Behalf Of Borries
> Demeler
> Sent: April 1, 2021 5:42 PM
> To: Myotoxin-II discussion <myotox at biophysics.uleth.ca>
> Subject: Re: [Myotox] myotoxin2-SDS titration
>
> Caution: This email was sent from someone outside of the University of
> Lethbridge. Do not click on links or open attachments unless you know they
> are safe. Suspicious emails should be forwarded to phishing at uleth.ca.
>
>
> Varying the speed only affects the resolution:
>
> high speed: good resolution on s, poor resolution on D low speed: good
> resolution on D, poor resolution on s.
>
> -Borries
>
> On Thu, Apr 01, 2021 at 11:16:34PM +0000, Paul Hazendonk wrote:
> > Can you do dynamic studies by varying the speed of centrifugation?  I.e.
> the slope as a function of speed?
> >
> > From: Myotox <myotox-bounces at biophysics.uleth.ca> On Behalf Of Borries
> > Demeler
> > Sent: April 1, 2021 4:37 PM
> > To: Myotoxin-II discussion <myotox at biophysics.uleth.ca>
> > Subject: Re: [Myotox] myotoxin2-SDS titration
> >
> > Caution: This email was sent from someone outside of the University of
> Lethbridge. Do not click on links or open attachments unless you know they
> are safe. Suspicious emails should be forwarded to phishing at uleth.ca
> <mailto:phishing at uleth.ca>.
> >
> > The s-value is reflective of the size of the molecule. A 5.5 s species
> is much bigger than a 2 s species. The y axis tells you the relative amount
> of species with a given s-value. It is a standard integral sedimentation
> distribution. I am happy to have a zoom meeting to discuss further these
> interesting trends.
> > -Borries
> >
> > On Thu, Apr 1, 2021 at 4:20 PM Hazendonk, Paul <paul.hazendonk at uleth.ca
> <mailto:paul.hazendonk at uleth.ca>> wrote:
> > What is the best way to interpret these curves.  I have not seen
> ultracentrifugation data since 1991.  Should we be looking at the position
> of the inflection point?
> >
> >
> > -----Original Message-----
> > From: Myotox
> > <myotox-bounces at biophysics.uleth.ca<mailto:myotox-bounces at biophysics.u
> > leth.ca>> On Behalf Of Borries Demeler
> > Sent: April 1, 2021 11:57 AM
> > To: Myotoxin-II discussion
> > <myotox at biophysics.uleth.ca<mailto:myotox at biophysics.uleth.ca>>
> > Subject: Re: [Myotox] myotoxin2-SDS titration
> >
> > Caution: This email was sent from someone outside of the University of
> Lethbridge. Do not click on links or open attachments unless you know they
> are safe. Suspicious emails should be forwarded to phishing at uleth.ca
> <mailto:phishing at uleth.ca>.
> >
> >
> > Dear all,
> > we have results back from the myotoxin-2 SDS titration experiments in
> AUC. I feel these experiments provided some very interesting results and
> are probably the most informative ones we have done so far. I also think
> that they will be extremely helpful in designing the *correct* NMR
> experiment. I went through the effort to write this up with figures, please
> review carefully the attached document and read the explanations I believe
> explain these results (which were quite unintuitive at first, but now make
> sense to me). Then let's schedule another meeting and discussion of next
> steps.
> >
> > -Borries
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