[3dem] [External] Re: Membrane protein dissociating from nanodisc

[Dykstra, Holly]

Yes, we use a mix of lipids.

From: AMARSHI MUKHERJEE <amarshimukherjee at gmail.com>
Sent: Friday, October 15, 2021 11:55 AM
To: Dykstra, Holly <Holly.Dykstra at vai.org>
Cc: 3dem at ncmir.ucsd.edu
Subject: [External] Re: [3dem] Membrane protein dissociating from nanodisc

Hi,

What is the lipid composition of the nanodiscs? If you are using a single lipid (such as PC only) the chance of incorporation of protein is low sometimes and may cause protein dissociation. if you use a mixture of lipids (such as PC/PS or PC/PE/PS) it may stabilize the protein in nanodiscs more than a single lipid nanodiscs.

On Fri, Oct 15, 2021 at 8:13 AM Dykstra, Holly <Holly.Dykstra at vai.org<mailto:Holly.Dykstra at vai.org>> wrote:
Hello all,
I am working on a single-TM membrane protein in nanodisc. Upon freezing grids, I am observing protein dissociation from the nanodisc. A small percentage of the total particle population is intact protein-nanodisc complex but most of the particles have dissociated. I believe this is happening at the point of freezing as negative stain shows the intact protein-nanodisc complex. Has anyone else experienced this problem or does anyone have any suggestions or advice?
Thanks!
-Holly
_______________________________________________
3dem mailing list
3dem at ncmir.ucsd.edu<mailto:3dem at ncmir.ucsd.edu>
https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem


--
Regards,

Amarshi Mukherjee, M.Sc., Ph.D.
Research Associate
Department of Neuroscience
Wisconsin Institute for Medical Research (WIMR)
1111 Highland Avenue, Madison, WI 53705
Mobile: 443-683-6757
Email: amukherjee29 at wisc.edu<mailto:amukherjee29 at wisc.edu>, amarshimukherjee at gmail.com<mailto:amarshimukherjee at gmail.com>

CAUTION: This email was sent from outside of the organization (amarshimukherjee at gmail.com<mailto:amarshimukherjee at gmail.com>). Do not click links or open attachments unless you recognize the sender and know the content is safe. If you have any questions, please contact ServiceDesk at vai.org<mailto:servicedesk at vai.org>.
-------------- next part --------------
An HTML attachment was scrubbed...
URL: <http://mail.ncmir.ucsd.edu/pipermail/3dem/attachments/20211015/1e837f79/attachment-0001.html>