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| Title | DM: a simple solution to suppress air-water interface interactions in cryo-EM. |
|---|---|
| Journal, issue, pages | bioRxiv, Year 2026 |
| Publish date | Apr 5, 2026 |
Authors | Maria Rafiq / Jan-Hannes Schäfer / Hamidreza Rahmani / Shaochen You / Michael Bollong / Danielle Grotjahn / R Luke Wiseman / Gabriel C Lander / ![]() |
| PubMed Abstract | The air-water interface (AWI) remains the primary barrier to routine high-resolution cryo-EM structure determination, driving protein adsorption, structural denaturation, and restricted particle ...The air-water interface (AWI) remains the primary barrier to routine high-resolution cryo-EM structure determination, driving protein adsorption, structural denaturation, and restricted particle orientations during vitrification. Here, we describe a simple and broadly applicable strategy to mitigate these effects using the mild non-ionic detergent n-decyl-β-D-maltopyranoside (DM). Addition of DM at low millimolar concentrations immediately prior to vitrification consistently suppresses AWI-driven artifacts, resulting in improved angular sampling, reduced structural damage, and enhanced reconstruction quality across diverse macromolecular systems. Using this approach, we obtained a high-resolution reconstruction of the 65 kDa Nucleophosmin 1 pentamer, a target previously limited by severe preferred orientation issues. We further show that DM promotes isotropic particle distributions for high-resolution reconstruction of hemagglutinin, transthyretin, as well as suppressing denaturation of aldolase while stabilizing its C-terminus. Our results indicate that DM effectively passivates deleterious air-water interface interactions without compromising particle integrity. These results establish DM as an effective additive for improving the robustness of single-particle cryo-EM sample preparation. |
External links | bioRxiv / PubMed:41959516 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 3.2 Å |
| Structure data | EMDB-74011, PDB-9zbv: |
| Source |
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Keywords | TRANSPORT PROTEIN / thyroxine transporter |
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