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TitleCryo-Electron Microscopy Snapshots of Eukaryotic Membrane Proteins in Native Lipid-Bilayer Nanodiscs.
Journal, issue, pagesBiomacromolecules, Vol. 23, Issue 12, Page 5084-5094, Year 2022
Publish dateDec 12, 2022
AuthorsKevin Janson / Fotis L Kyrilis / Christian Tüting / Marie Alfes / Manabendra Das / Toni K Träger / Carla Schmidt / Farzad Hamdi / Carolyn Vargas / Sandro Keller / Annette Meister / Panagiotis L Kastritis /
PubMed AbstractNew technologies for purifying membrane-bound protein complexes in combination with cryo-electron microscopy (EM) have recently allowed the exploration of such complexes under near-native conditions. ...New technologies for purifying membrane-bound protein complexes in combination with cryo-electron microscopy (EM) have recently allowed the exploration of such complexes under near-native conditions. In particular, polymer-encapsulated nanodiscs enable the study of membrane proteins at high resolution while retaining protein-protein and protein-lipid interactions within a lipid bilayer. However, this powerful technology has not been exploited to address the important question of how endogenous─as opposed to overexpressed─membrane proteins are organized within a lipid environment. In this work, we demonstrate that biochemical enrichment protocols for native membrane-protein complexes from in combination with polymer-based lipid-bilayer nanodiscs provide a substantial improvement in the quality of recovered endogenous membrane-protein complexes. Mass spectrometry results revealed ∼1123 proteins, while multiple 2D class averages and two 3D reconstructions from cryo-EM data furnished prominent structural signatures. This integrated methodological approach to enriching endogenous membrane-protein complexes provides unprecedented opportunities for a deeper understanding of eukaryotic membrane proteomes.
External linksBiomacromolecules / PubMed:36399657
MethodsEM (single particle)
Resolution4.73 - 18.59 Å
Structure data

EMDB-15516: Cryo-EM Snapshots of Nanodisc-Embedded Native Eukaryotic Membrane Proteins
Method: EM (single particle) / Resolution: 18.59 Å

EMDB-15517: myo-Inositol-1-Phosphate Synthase
Method: EM (single particle) / Resolution: 4.73 Å

Source
  • Thermochaetoides thermophila (fungus)

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