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<p class="MsoNormal">Tim,<o:p></o:p></p>
<p class="MsoNormal">We use MSP1E3. Have you embedded single TM in SMALP and seen the protein stay in intact?<o:p></o:p></p>
<p class="MsoNormal">-Holly <o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal"><b>From:</b> Timothy Dafforn <T.R.Dafforn@bham.ac.uk> <br>
<b>Sent:</b> Friday, October 15, 2021 10:27 AM<br>
<b>To:</b> jacopo.marino@psi.ch; Dykstra, Holly <Holly.Dykstra@vai.org><br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> [External] Re: [3dem] Membrane protein dissociating from nanodisc<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p><span style="font-size:12.0pt;color:black">what form of nano disc are you using?<o:p></o:p></span></p>
<p><span style="font-size:12.0pt;color:black">We do a fair amount of work on the SMALP system (we developed it) so might be able to help<o:p></o:p></span></p>
<p><span style="font-size:12.0pt;color:black">Tim<o:p></o:p></span></p>
<p><span style="font-size:12.0pt;color:black"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black"><br>
Tim R. Dafforn PhD FIET<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size:10.0pt;font-family:"Tahoma",sans-serif;color:black">Professor of Biotechnology<br>
University of Birmingham<br>
Edgbaston<br>
Birmingham<br>
B152TT<br>
+44 (0) 121 414 5881<o:p></o:p></span></p>
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<p class="MsoNormal"><b><span style="color:black">From:</span></b><span style="color:black"> 3dem <<a href="mailto:3dem-bounces@ncmir.ucsd.edu">3dem-bounces@ncmir.ucsd.edu</a>> on behalf of
<a href="mailto:jacopo.marino@psi.ch">jacopo.marino@psi.ch</a> <<a href="mailto:jacopo.marino@psi.ch">jacopo.marino@psi.ch</a>><br>
<b>Sent:</b> 15 October 2021 14:39:07<br>
<b>To:</b> Dykstra, Holly<br>
<b>Cc:</b> <a href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a><br>
<b>Subject:</b> Re: [3dem] Membrane protein dissociating from nanodisc</span> <o:p>
</o:p></p>
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<p class="MsoNormal"> <o:p></o:p></p>
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<p class="MsoNormal">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.
<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal">Cheers<o:p></o:p></p>
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<p class="MsoNormal" style="margin-bottom:12.0pt">Jacopo <o:p></o:p></p>
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<pre><span style="font-family:".SFUIDisplay",serif">Dr. Jacopo Marino Laboratory of Biomolecular Research Paul Scherrer Institute OFLB/005 5232 </span><o:p></o:p></pre>
<pre><span style="font-family:".SFUIDisplay",serif">Villigen PSI, Switzerland</span><o:p></o:p></pre>
<pre><span style="font-family:".SFUIDisplay",serif">tel: <a href="tel:+41%2056%20310%205777">+41 56 310 5777</a></span><o:p></o:p></pre>
<pre><span style="font-family:".SFUIDisplay",serif">e-mail: <a href="mailto:jacopo.marino@psi.ch">jacopo.marino@psi.ch</a></span><o:p></o:p></pre>
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On 15 Oct 2021, at 15:13, Dykstra, Holly <<a href="mailto:Holly.Dykstra@vai.org">Holly.Dykstra@vai.org</a>> wrote:<o:p></o:p></p>
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<p class="MsoNormal">Hello all,<o:p></o:p></p>
<p class="MsoNormal">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?<o:p></o:p></p>
<p class="MsoNormal">Thanks!<o:p></o:p></p>
<p class="MsoNormal">-Holly<o:p></o:p></p>
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<p style="background:#FFEB9C"><strong><span style="font-size:10.0pt;font-family:"Calibri",sans-serif;color:#9C6500;background:#FFEB9C">CAUTION</span></strong><strong><span style="font-size:8.5pt;font-family:"yui-tmp",serif;color:#9C6500;background:#FFEB9C">:</span></strong><span style="font-size:10.0pt;color:black"> This
email was sent from outside of the organization (<a href="mailto:T.R.Dafforn@bham.ac.uk">T.R.Dafforn@bham.ac.uk</a>). 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
<a href="mailto:servicedesk@vai.org">ServiceDesk@vai.org</a>.</span><o:p></o:p></p>
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