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<p class="MsoNormal"><span style="mso-fareast-language:EN-US">I though maybe that the width of the band would be functionally dependent on the reaction rate, binding mechanism, and diffusion constant.
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<p class="MsoNormal"><b><span lang="EN-US">From:</span></b><span lang="EN-US"> Myotox <myotox-bounces@biophysics.uleth.ca>
<b>On Behalf Of </b>Borries Demeler<br>
<b>Sent:</b> April 1, 2021 10:01 PM<br>
<b>To:</b> Myotoxin-II discussion <myotox@biophysics.uleth.ca><br>
<b>Subject:</b> Re: [Myotox] myotoxin2-SDS titration<o:p></o:p></span></p>
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<span style="font-size:10.0pt;color:black">Caution: This email was sent from someone
<b>outside of the University of Lethbridge</b>. Do not click on links or open attachments unless you know they are safe. Suspicious emails should be forwarded to
<a href="mailto:phishing@uleth.ca">phishing@uleth.ca</a>.</span><o:p></o:p></p>
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<p class="MsoNormal">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.
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<p class="MsoNormal">-b.<o:p></o:p></p>
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<p class="MsoNormal">On Thu, Apr 1, 2021 at 6:16 PM Hazendonk, Paul <<a href="mailto:paul.hazendonk@uleth.ca">paul.hazendonk@uleth.ca</a>> wrote:<o:p></o:p></p>
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<p class="MsoNormal">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.<br>
<br>
-----Original Message-----<br>
From: Myotox <<a href="mailto:myotox-bounces@biophysics.uleth.ca" target="_blank">myotox-bounces@biophysics.uleth.ca</a>> On Behalf Of Borries Demeler<br>
Sent: April 1, 2021 5:42 PM<br>
To: Myotoxin-II discussion <<a href="mailto:myotox@biophysics.uleth.ca" target="_blank">myotox@biophysics.uleth.ca</a>><br>
Subject: Re: [Myotox] myotoxin2-SDS titration<br>
<br>
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
<a href="mailto:phishing@uleth.ca" target="_blank">phishing@uleth.ca</a>.<br>
<br>
<br>
Varying the speed only affects the resolution:<br>
<br>
high speed: good resolution on s, poor resolution on D low speed: good resolution on D, poor resolution on s.<br>
<br>
-Borries<br>
<br>
On Thu, Apr 01, 2021 at 11:16:34PM +0000, Paul Hazendonk wrote:<br>
> Can you do dynamic studies by varying the speed of centrifugation? I.e. the slope as a function of speed?<br>
><br>
> From: Myotox <<a href="mailto:myotox-bounces@biophysics.uleth.ca" target="_blank">myotox-bounces@biophysics.uleth.ca</a>> On Behalf Of Borries
<br>
> Demeler<br>
> Sent: April 1, 2021 4:37 PM<br>
> To: Myotoxin-II discussion <<a href="mailto:myotox@biophysics.uleth.ca" target="_blank">myotox@biophysics.uleth.ca</a>><br>
> Subject: Re: [Myotox] myotoxin2-SDS titration<br>
><br>
> 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
<a href="mailto:phishing@uleth.ca" target="_blank">phishing@uleth.ca</a><mailto:<a href="mailto:phishing@uleth.ca" target="_blank">phishing@uleth.ca</a>>.<br>
><br>
> 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.<br>
> -Borries<br>
><br>
> On Thu, Apr 1, 2021 at 4:20 PM Hazendonk, Paul <<a href="mailto:paul.hazendonk@uleth.ca" target="_blank">paul.hazendonk@uleth.ca</a><mailto:<a href="mailto:paul.hazendonk@uleth.ca" target="_blank">paul.hazendonk@uleth.ca</a>>> wrote:<br>
> 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?<br>
><br>
><br>
> -----Original Message-----<br>
> From: Myotox <br>
> <<a href="mailto:myotox-bounces@biophysics.uleth.ca" target="_blank">myotox-bounces@biophysics.uleth.ca</a><mailto:<a href="mailto:myotox-bounces@biophysics.u" target="_blank">myotox-bounces@biophysics.u</a><br>
> <a href="http://leth.ca" target="_blank">leth.ca</a>>> On Behalf Of Borries Demeler<br>
> Sent: April 1, 2021 11:57 AM<br>
> To: Myotoxin-II discussion <br>
> <<a href="mailto:myotox@biophysics.uleth.ca" target="_blank">myotox@biophysics.uleth.ca</a><mailto:<a href="mailto:myotox@biophysics.uleth.ca" target="_blank">myotox@biophysics.uleth.ca</a>>><br>
> Subject: Re: [Myotox] myotoxin2-SDS titration<br>
><br>
> 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
<a href="mailto:phishing@uleth.ca" target="_blank">phishing@uleth.ca</a><mailto:<a href="mailto:phishing@uleth.ca" target="_blank">phishing@uleth.ca</a>>.<br>
><br>
><br>
> Dear all,<br>
> 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.<br>
><br>
> -Borries<br>
> _______________________________________________<br>
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> <a href="https://demeler7.uleth.ca/mailman/listinfo/myotox" target="_blank">https://demeler7.uleth.ca/mailman/listinfo/myotox</a><br>
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