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<p>Dear colleagues</p>
<p><br>
</p>
<p>thanks for running these new experiments, and finding such
interesting results!</p>
<p><br>
</p>
<p>so this dimerization induced by SDS would explain the appearance
of a smear around the dimeric value in such electrophoretic
analyses? and this would support that under physiological
conditions the toxin would be a monomer, the dimer being an
artifact of crystallization perhaps? I am not sure if I am
interpreting this correctly</p>
<p><br>
</p>
<p>exciting news indeed</p>
<p><br>
</p>
<p>very best regards and thank you so much<br>
</p>
<p><br>
</p>
<p>Bruno</p>
<p><br>
</p>
<p><br>
</p>
<p><br>
</p>
<p>PS: it is likely that I will have an important surgery in the
next days, which might keep me a bit disconnected for few weeks,
in case you do not see me replying quickly to mails</p>
<p><br>
</p>
<p><br>
</p>
<p>+++++</p>
<p><br>
</p>
<p><br>
</p>
<p><br>
</p>
<div class="moz-cite-prefix">On 3/12/2020 4:52 PM, Henrickson, Amy
wrote:<br>
</div>
<blockquote type="cite"
cite="mid:YQXPR0101MB1910CAC7339FC96EBCA59079EFFD0@YQXPR0101MB1910.CANPRD01.PROD.OUTLOOK.COM">
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Hello,</div>
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;
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<br>
</div>
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;
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Just to clarify the SDS concentration was 1%(w/v). <br>
</div>
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;
font-size:12pt; color:rgb(0,0,0)">
<br>
</div>
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;
font-size:12pt; color:rgb(0,0,0)">
Cheers, <br>
</div>
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;
font-size:12pt; color:rgb(0,0,0)">
Amy Henrickson <br>
</div>
<hr tabindex="-1" style="display:inline-block; width:98%">
<div id="divRplyFwdMsg" dir="ltr"><font style="font-size:11pt"
face="Calibri, sans-serif" color="#000000"><b>From:</b> Myotox
<a class="moz-txt-link-rfc2396E" href="mailto:myotox-bounces@biophysics.uleth.ca"><myotox-bounces@biophysics.uleth.ca></a> on behalf of
Borries Demeler <a class="moz-txt-link-rfc2396E" href="mailto:demeler@gmail.com"><demeler@gmail.com></a><br>
<b>Sent:</b> Thursday, March 12, 2020 2:45 PM<br>
<b>To:</b> <a class="moz-txt-link-abbreviated" href="mailto:myotox@biophysics.uleth.ca">myotox@biophysics.uleth.ca</a>
<a class="moz-txt-link-rfc2396E" href="mailto:myotox@biophysics.uleth.ca"><myotox@biophysics.uleth.ca></a><br>
<b>Subject:</b> [Myotox] SDS AUC update</font>
<div> </div>
</div>
<div class="BodyFragment"><font size="2"><span
style="font-size:11pt">
<div class="PlainText">Dear all,<br>
Amy ran the most recent samples from Bruno in the presence
of 1 mg/ml<br>
SDS at two protein concentrations (1 mg/ml and 0.17
mg/ml). The results<br>
are quite consistent with the results in Susumu's paper.
We do see<br>
oligomerization, with a more or less final state. At this
point I am not<br>
clear if it is a dimer or trimer or something higher, this
still needs<br>
to be determined. But it is very clear that SDS has a big
effect on<br>
dimerization. What also is interesting is that the
association behavior<br>
induced by SDS appears to be reversible. That is, you get
dissociation<br>
of the oligomer in response to lowering the concentration.
We will<br>
further test this by using even lower concentration to see
if we can<br>
shift to entirely monomer. We will also do D2O density
matching to see<br>
if there is a density difference between monomer and dimer
and<br>
potentially higher oligomers due to different ratios of
SDS binding to<br>
myotoxin-II.<br>
<br>
Now we need to come up with a strutural explanation why
this<br>
oligomerization shift is happening in response to addition
of SDS. I<br>
would also like to know if some other lipids could cause
the same effect<br>
as a detergent? A figure is attached that shows the
sedimentation<br>
distribution, it looks like similar values as those
obtained for the<br>
different toxin in the Uchiyama paper.<br>
<br>
Regards, -Borries<br>
<br>
<br>
</div>
</span></font></div>
<br>
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<pre class="moz-quote-pre" wrap="">_______________________________________________
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</pre>
</blockquote>
<pre class="moz-signature" cols="72">--
Bruno Lomonte, Ph.D.
Instituto Clodomiro Picado
Universidad de Costa Rica
San José, 11501
COSTA RICA
<a class="moz-txt-link-abbreviated" href="mailto:bruno.lomonte@ucr.ac.cr">bruno.lomonte@ucr.ac.cr</a>
tel. +506 2511 7888
cel. +506 8392 0012
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