[3dem] Membrane protein dissociating from nanodisc

[Jacopo Marino]

Hi - one would expect that membrane proteins embedded into nanodiscs would suffer less than detergent-embedded proteins, but that’s not a rule. The question is simple: is it faster to live with your small amount of particles and compensate with collecting many more datasets, or screening for freezing conditions where you have less dissociation ? You might find that areas with thicker ice on your grid have more intact particles, but likely you don’t want to collect from there. 

Cheers
Jacopo 

Dr. Jacopo Marino Laboratory of Biomolecular Research Paul Scherrer Institute OFLB/005 5232 
Villigen PSI, Switzerland
tel: +41 56 310 5777
e-mail: jacopo.marino at psi.ch

> On 15 Oct 2021, at 15:13, Dykstra, Holly <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
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