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TitleDM: a simple solution to suppress air-water interface interactions in cryo-EM.
Journal, issue, pagesbioRxiv, Year 2026
Publish dateApr 5, 2026
AuthorsMaria Rafiq / Jan-Hannes Schäfer / Hamidreza Rahmani / Shaochen You / Michael Bollong / Danielle Grotjahn / R Luke Wiseman / Gabriel C Lander /
PubMed AbstractThe 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 linksbioRxiv / PubMed:41959516 / PubMed Central
MethodsEM (single particle)
Resolution3.2 Å
Structure data

EMDB-74011, PDB-9zbv:
Human TTR-C10A at pH 7.4
Method: EM (single particle) / Resolution: 3.2 Å

Source
  • homo sapiens (human)
KeywordsTRANSPORT PROTEIN / thyroxine transporter

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