[Myotox] [tony.montina at uleth.ca: RE: Myotoxin-II]

[Henrickson, Amy]

Hello,

For the NMR studies I provided about 2mg of sample therefore they would like 4-6mg more.

I do still have what I would guess to be about 0.5mg of lyophilized sample left.
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.

Cheers,

Amy Henrickson
________________________________
From: Myotox <myotox-bounces at biophysics.uleth.ca> on behalf of Borries Demeler <demeler at gmail.com>
Sent: Sunday, June 7, 2020 12:31 PM
To: myotox at biophysics.uleth.ca <myotox at biophysics.uleth.ca>
Subject: [Myotox] [tony.montina at uleth.ca: RE: Myotoxin-II]

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 phishing at uleth.ca.

Hello Bruno,
Paul's group completed the first NMR experiment on Myotoxin-II, which
according to Paul and Tony provided very nice signal. I thought it
would be useful to compare the +/- SDS conditions by NMR as well and
see if we can learn something new about the interaction of SDS with
myotoxin-II. Everybody, please let us know if you agree with this.

As you can see from the communication below, Tony would require additional
protein to perform these studies and run the necessary controls. Amy,
can you please advise Bruno of the amounts? It would be 2-3 times the
amount you gave to Tony previously. Once we know the amounts, please
let us know if it is possible to send one more batch of myotoxin-II.

My guess is that it would be best if we could obtain the sample in
lyophilized format, so it can be resuspended directly in aqueous buffer
made with either light or heavy water. Importantly, if the protein is
dissolved in D2O, lyophilized sample would allow us to reconstitute
directly in D2O and avoid contamination by light water, which gives
different background signals in NMR. Tony, please confirm if this is
what you would like to have. THe SDS concentration should be 1%, which
is what we measured by AUC.

Paul: Can you please describe the results you obtained and let us know
based on the preliminary data what we can expect to learn from the
additional NMR experiments, especially with respect to protein structure
and interaction with SDS. If this provides greater insights into
the strange protein behavior in SDS I think it would really make our
manuscript a lot more interesting!

Thank you all! -Borries


----- Forwarded message from "Montina, Tony" <tony.montina at uleth.ca> -----

Date: Sun, 7 Jun 2020 18:04:01 +0000
From: "Montina, Tony" <tony.montina at uleth.ca>
To: Borries Demeler <demeler at gmail.com>, "Hazendonk, Paul" <paul.hazendonk at uleth.ca>
CC: "Henrickson, Amy" <amy.henrickson at uleth.ca>
Subject: RE: Myotoxin-II

Hello Borries,
I believe that Mike used most of the sample that Amy gave him to make up the myotoxin in D2O and PBS.
It would probably be worthwhile to ask Bruno to send more, as we will want to prepare the NMR samples under several conditions.
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.
Amy would have a better idea of how much she just gave us, but I would suggest two to three times that amount.
Does that work
Tony

_______________________________________________
Tony Montina
Director, Science Operations
Director, Magnetic Resonance Facility
Instructor, Department of Chemistry and Biochemistry
Faculty of Arts and Science
The University of Lethbridge
Lethbridge, Alberta, Canada, T1K 3M4
Office: 1-403-394-3927
Lab: 1-403-329-2230

-----Original Message-----
From: Borries Demeler <demeler at gmail.com>
Sent: June 5, 2020 4:23 PM
To: Hazendonk, Paul <paul.hazendonk at uleth.ca>
Cc: Montina, Tony <tony.montina at uleth.ca>; Henrickson, Amy <amy.henrickson at uleth.ca>
Subject: Re: Myotoxin-II

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 phishing at uleth.ca.

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.


On Fri, Jun 05, 2020 at 07:02:11PM +0000, Hazendonk, Paul wrote:
> Hi Borries
>
> 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.
>
> Cheers
>
> Paul
>
> Sent from Outlook Mobile<https://aka.ms/blhgte>
>

----- End forwarded message -----
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