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<p>Hello all,</p>
<p><br>
</p>
<p>very happy to hear that the NMR experiment went OK! and agree
that it would be nice to complement results with additional
comparisons in the presence of SDS or other conditions<br>
</p>
<p><br>
</p>
<p>I will prepare more myotoxin-II, although it may take a bit
longer than usual (since I have limited access to the institute
due to the coronavirus confinement). In any case I will be in
contact when ready to ship.</p>
<p><br>
</p>
<p>Hope you are all fine! here the number of cases was going really
down - good work done by the goverment - , but in these last days
things went out of hands with our neighbors in the North
(Nicaragua), desperately crossing the border illegally (and I
fully understand their situation) and now infections are again
growing in Costa Rica:</p>
<p><br>
</p>
<p><img src="cid:part1.B0856F2E.58864ABE@ucr.ac.cr" alt=""
width="503" height="296"></p>
<p><br>
</p>
<p><img src="cid:part2.AE492720.B942AB5E@ucr.ac.cr" alt=""
width="165" height="80"> <img
src="cid:part3.AACE4A92.53FD0B78@ucr.ac.cr" alt="" width="116"
height="83"></p>
<p><br>
</p>
<p>Best wishes to all,</p>
<p><br>
</p>
<p>Bruno<br>
</p>
<p><br>
</p>
<p><br>
</p>
<p>+++<br>
</p>
<p><br>
</p>
<p><br>
</p>
<div class="moz-cite-prefix">On 6/7/2020 3:05 PM, Henrickson, Amy
wrote:<br>
</div>
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Hello,</div>
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<br>
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For the NMR studies I provided about 2mg of sample therefore
they would like 4-6mg more.
<br>
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<br>
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font-size: 12pt; color: rgb(0, 0, 0);">
I do still have what I would guess to be about 0.5mg of
lyophilized sample left. <br>
</div>
<div style="font-family: Calibri, Arial, Helvetica, sans-serif;
font-size: 12pt; color: rgb(0, 0, 0);">
I also saved most of the samples that I ran in the AUC however,
they have been sitting at room temp suspended in buffer,
therefore I am not sure if it would be useful or not.
<br>
</div>
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<br>
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Cheers, <br>
</div>
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<br>
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<div style="font-family: Calibri, Arial, Helvetica, sans-serif;
font-size: 12pt; color: rgb(0, 0, 0);">
Amy Henrickson <br>
</div>
<hr style="display:inline-block;width:98%" tabindex="-1">
<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> Sunday, June 7, 2020 12:31 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] [<a class="moz-txt-link-abbreviated" href="mailto:tony.montina@uleth.ca">tony.montina@uleth.ca</a>: RE:
Myotoxin-II]</font>
<div> </div>
</div>
<div class="BodyFragment"><font size="2"><span
style="font-size:11pt;">
<div class="PlainText">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. Please forward suspicious emails to
<a class="moz-txt-link-abbreviated" href="mailto:phishing@uleth.ca">phishing@uleth.ca</a>.<br>
<br>
Hello Bruno,<br>
Paul's group completed the first NMR experiment on
Myotoxin-II, which<br>
according to Paul and Tony provided very nice signal. I
thought it<br>
would be useful to compare the +/- SDS conditions by NMR
as well and<br>
see if we can learn something new about the interaction of
SDS with<br>
myotoxin-II. Everybody, please let us know if you agree
with this.<br>
<br>
As you can see from the communication below, Tony would
require additional<br>
protein to perform these studies and run the necessary
controls. Amy,<br>
can you please advise Bruno of the amounts? It would be
2-3 times the<br>
amount you gave to Tony previously. Once we know the
amounts, please<br>
let us know if it is possible to send one more batch of
myotoxin-II.<br>
<br>
My guess is that it would be best if we could obtain the
sample in<br>
lyophilized format, so it can be resuspended directly in
aqueous buffer<br>
made with either light or heavy water. Importantly, if the
protein is<br>
dissolved in D2O, lyophilized sample would allow us to
reconstitute<br>
directly in D2O and avoid contamination by light water,
which gives<br>
different background signals in NMR. Tony, please confirm
if this is<br>
what you would like to have. THe SDS concentration should
be 1%, which<br>
is what we measured by AUC.<br>
<br>
Paul: Can you please describe the results you obtained and
let us know<br>
based on the preliminary data what we can expect to learn
from the<br>
additional NMR experiments, especially with respect to
protein structure<br>
and interaction with SDS. If this provides greater
insights into<br>
the strange protein behavior in SDS I think it would
really make our<br>
manuscript a lot more interesting!<br>
<br>
Thank you all! -Borries<br>
<br>
<br>
----- Forwarded message from "Montina, Tony"
<a class="moz-txt-link-rfc2396E" href="mailto:tony.montina@uleth.ca"><tony.montina@uleth.ca></a> -----<br>
<br>
Date: Sun, 7 Jun 2020 18:04:01 +0000<br>
From: "Montina, Tony" <a class="moz-txt-link-rfc2396E" href="mailto:tony.montina@uleth.ca"><tony.montina@uleth.ca></a><br>
To: Borries Demeler <a class="moz-txt-link-rfc2396E" href="mailto:demeler@gmail.com"><demeler@gmail.com></a>, "Hazendonk,
Paul" <a class="moz-txt-link-rfc2396E" href="mailto:paul.hazendonk@uleth.ca"><paul.hazendonk@uleth.ca></a><br>
CC: "Henrickson, Amy" <a class="moz-txt-link-rfc2396E" href="mailto:amy.henrickson@uleth.ca"><amy.henrickson@uleth.ca></a><br>
Subject: RE: Myotoxin-II<br>
<br>
Hello Borries,<br>
I believe that Mike used most of the sample that Amy gave
him to make up the myotoxin in D2O and PBS.<br>
It would probably be worthwhile to ask Bruno to send more,
as we will want to prepare the NMR samples under several
conditions.<br>
For example, we might want a sample (as you suggested)
with SDS, we might want another sample with H2O and D2O
(for exchange purposes), etc.<br>
Amy would have a better idea of how much she just gave us,
but I would suggest two to three times that amount.<br>
Does that work<br>
Tony<br>
<br>
_______________________________________________<br>
Tony Montina<br>
Director, Science Operations<br>
Director, Magnetic Resonance Facility<br>
Instructor, Department of Chemistry and Biochemistry<br>
Faculty of Arts and Science<br>
The University of Lethbridge<br>
Lethbridge, Alberta, Canada, T1K 3M4<br>
Office: 1-403-394-3927<br>
Lab: 1-403-329-2230<br>
<br>
-----Original Message-----<br>
From: Borries Demeler <a class="moz-txt-link-rfc2396E" href="mailto:demeler@gmail.com"><demeler@gmail.com></a><br>
Sent: June 5, 2020 4:23 PM<br>
To: Hazendonk, Paul <a class="moz-txt-link-rfc2396E" href="mailto:paul.hazendonk@uleth.ca"><paul.hazendonk@uleth.ca></a><br>
Cc: Montina, Tony <a class="moz-txt-link-rfc2396E" href="mailto:tony.montina@uleth.ca"><tony.montina@uleth.ca></a>;
Henrickson, Amy <a class="moz-txt-link-rfc2396E" href="mailto:amy.henrickson@uleth.ca"><amy.henrickson@uleth.ca></a><br>
Subject: Re: Myotoxin-II<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. Please forward
suspicious emails to <a class="moz-txt-link-abbreviated" href="mailto:phishing@uleth.ca">phishing@uleth.ca</a>.<br>
<br>
Tony/Amy, Paul mentioned that we could do this experiment
in the presence of SDS. How much material would we need
and how much do we still have on hand? If we don't have
enough we should contact Bruno and ask him to milk some
more snakes :) -b.<br>
<br>
<br>
On Fri, Jun 05, 2020 at 07:02:11PM +0000, Hazendonk, Paul
wrote:<br>
> Hi Borries<br>
><br>
> We have a 1D proton that makes sense to me now. We
have some T1 data. I am waiting for the T2 and DOSY
results. Mike could you updaye us on when these will be
done.<br>
><br>
> Cheers<br>
><br>
> Paul<br>
><br>
> Sent from Outlook Mobile<<a
href="https://aka.ms/blhgte" moz-do-not-send="true">https://aka.ms/blhgte</a>><br>
><br>
<br>
----- End forwarded message -----<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
</pre>
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