[Myotox] Myotoxin work

[Borries Demeler]

Hi Tony,
> Hello Amy and Borries,
> Thank you for the update.
> I apologize for not getting back to either of your last week, but the week was very busy.
> I think we should make a more concentrated sample now that you have more material. Even 1mg/mL is not very concentrated.
> Can we make a more concentrated sample with the new material?

Yes, I think we have enough material to make a more concentrated sample.
Please be specific about volume and concentration, and we'll try to make
it.

> I also think this would be best to run the NMR on this sample at analogous buffer conditions to the AUC experiments, so long as it is not too lossy (salty).

Regarding buffer:
We already have AUC data performed in PBS, but we can rerun in any other
buffer that you would like to test for NMR. So please let us know what
would be optimal. We do like to have an ionic strength of ~ 25 mM, 5-10
mM NaCl would be desirable at a minimum. The rest could be buffer,
either TRIS or phosphate, let us know what you want.

Right now we are testing myotoxin-II in the presence of SDS to see if
there is an effect when aliphatics are around and may induce
dimerization. Later, we may switch to lipid bilayers in nanodiscs.

> Paul, I was thinking to start with the basic 1D and 2D experiments (COSY, TOCSY, HSQC, HMBC), just like we did with the T2 toxin in the past.
> Could all of us arrange a meeting sometime soon so that we can finalize the details of the sample, the buffer to use, and also the exact NMR experiments we would like to start with?

I am around all week and fairly flexible, except tomorrow.

Regards, -Borries

> Please let me know if everyone wants to meet or not.
> Cheers
> Tony
> 
> Tony Montina
> Interim Director, Science Commons Academic Operations
> Director, Magnetic Resonance Facility
> Instructor, Department of Chemistry and Biochemistry
> The University of Lethbridge,
> 4401 University Drive West
> Lethbridge, Alberta, Canada, T1K 3M4
> Office: SA6214 Science Commons 1-403-394-3927
> Lab: SA6216 Science Commons 1-403-329-2230
> 
> From: Henrickson, Amy
> Sent: February 14, 2020 11:49 AM
> To: Montina, Tony <tony.montina at uleth.ca>; Hazendonk, Paul <paul.hazendonk at uleth.ca>
> Cc: Opyr, Michael <michael.opyr at uleth.ca>
> Subject: Re: Myotoxin work
> 
> Hello All,
> 
> I was wondering if you had had a chance to run this samples and if so how it turned out.
> 
> I have also received new samples from the collaborator, it is currently stored as a dried sample. I would be able to bring some down to you if you would like. I can also resuspend it in buffer for you if you would like.
> 
> Thanks,
> 
> Amy
> ________________________________
> From: Henrickson, Amy <amy.henrickson at uleth.ca<mailto:amy.henrickson at uleth.ca>>
> Sent: Friday, January 17, 2020 9:20 AM
> To: Montina, Tony <tony.montina at uleth.ca<mailto:tony.montina at uleth.ca>>; Hazendonk, Paul <paul.hazendonk at uleth.ca<mailto:paul.hazendonk at uleth.ca>>
> Cc: Opyr, Michael <michael.opyr at uleth.ca<mailto:michael.opyr at uleth.ca>>
> Subject: Re: Myotoxin work
> 
> Thank you Tomy.
> 
> Mike, would you be free for me to come down between 10-11? At this time I can bring the sample and all of the buffer/sample details as well.
> 
> Thanks,
> Amy
> 
> ________________________________
> From: Montina, Tony <tony.montina at uleth.ca<mailto:tony.montina at uleth.ca>>
> Sent: Thursday, January 16, 2020 7:07 AM
> To: Henrickson, Amy <amy.henrickson at uleth.ca<mailto:amy.henrickson at uleth.ca>>; Hazendonk, Paul <paul.hazendonk at uleth.ca<mailto:paul.hazendonk at uleth.ca>>
> Cc: Opyr, Michael <michael.opyr at uleth.ca<mailto:michael.opyr at uleth.ca>>
> Subject: RE: Myotoxin work
> 
> 
> Hello Amy,
> 
> I have made use of the lyophilizer in Steve Mosimann's lab in the past.
> 
> Maybe just bring the sample down to Mike and I in the MR Facility (6th level of the Science Commons) and we can get make arrangements to get it lyophilized.
> 
> When you bring the sample can you please bring the relevant sample information.
> 
> This would include all details on the buffer that the sample is currently in, the concentration of the buffer and sample, and the approximate volume.
> 
> I have cc'd Mike on this email so that you can make arrangements with him to drop off the sample.
> 
> Cheers
> Tony
> 
> 
> 
> Tony Montina
> 
> Interim Director, Science Commons Academic Operations
> 
> Director, Magnetic Resonance Facility
> 
> Instructor, Department of Chemistry and Biochemistry
> 
> The University of Lethbridge,
> 
> 4401 University Drive West
> 
> Lethbridge, Alberta, Canada, T1K 3M4
> 
> Office: SA6214 Science Commons 1-403-394-3927
> 
> Lab: SA6216 Science Commons 1-403-329-2230
> 
> 
> 
> From: Henrickson, Amy
> Sent: January 15, 2020 1:34 PM
> To: Hazendonk, Paul <paul.hazendonk at uleth.ca<mailto:paul.hazendonk at uleth.ca>>; Montina, Tony <tony.montina at uleth.ca<mailto:tony.montina at uleth.ca>>
> Subject: Re: Myotoxin work
> 
> 
> 
> Hello Tony,
> 
> 
> 
> My name is Amy Henrickson and I am part of Borries Demeler's lab. We have been talking with Paul about running some snake venom proteins on the NMR, I was wondering if you had a lyophilizer that we could use to dry the sample so that it can be resuspended in D2O?
> 
> 
> 
> Thank you,
> 
> 
> 
> Amy Henrickson
> 
> ________________________________
> 
> From: Hazendonk, Paul <paul.hazendonk at uleth.ca<mailto:paul.hazendonk at uleth.ca>>
> Sent: Tuesday, January 14, 2020 1:43 PM
> To: Henrickson, Amy <amy.henrickson at uleth.ca<mailto:amy.henrickson at uleth.ca>>
> Subject: Re: Myotoxin work
> 
> 
> 
> Maybe Tony has access to one.
> 
> Get Outlook for Android<https://aka.ms/ghei36>
> 
> 
> 
> ________________________________
> 
> From: Henrickson, Amy <amy.henrickson at uleth.ca<mailto:amy.henrickson at uleth.ca>>
> Sent: Tuesday, January 14, 2020 10:54:56 AM
> To: Hazendonk, Paul <paul.hazendonk at uleth.ca<mailto:paul.hazendonk at uleth.ca>>
> Subject: Myotoxin work
> 
> 
> 
> Hello Paul,
> 
> 
> 
> This is Amy Henrickson, Borries Demeler's lab manager.
> 
> I was wondering if you had access to a lyophilizer so that we would be able to dry down the snake venom protein to redissolve in D2O, for your experiments?
> 
> 
> 
> Thanks,
> 
> Amy Henrickson
> 
> 
> 
> ---
> 
> Amy Henrickson B.Sc
> 
> Laboratory Manager Canadian Center for Hydrodynamics
> 
> Northwest Biophysics Consortium
> 
> Alberta RNA Research and Training Institute
> Department of Chemistry and Biochemistry
> University of Lethbridge
> 4401 University Drive
> Lethbridge, Alberta, Canada T1K 3M4
> 
> Email: amy.henrickson at uleth.ca<mailto:amy.henrickson at uleth.ca>
> 
> Phone: 403-329-2095
> 
> 
> 
> ----- End forwarded message -----