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TitleCryo-electron tomography pipeline for plasma membranes.
Journal, issue, pagesNat Commun, Vol. 16, Issue 1, Page 855, Year 2025
Publish dateJan 20, 2025
AuthorsWilly W Sun / Dennis J Michalak / Kem A Sochacki / Prasanthi Kunamaneni / Marco A Alfonzo-Méndez / Andreas M Arnold / Marie-Paule Strub / Jenny E Hinshaw / Justin W Taraska /
PubMed AbstractCryo-electron tomography (cryoET) provides sub-nanometer protein structure within the dense cellular environment. Existing sample preparation methods are insufficient at accessing the plasma membrane ...Cryo-electron tomography (cryoET) provides sub-nanometer protein structure within the dense cellular environment. Existing sample preparation methods are insufficient at accessing the plasma membrane and its associated proteins. Here, we present a correlative cryo-electron tomography pipeline optimally suited to image large ultra-thin areas of isolated basal and apical plasma membranes. The pipeline allows for angstrom-scale structure determination with subtomogram averaging and employs a genetically encodable rapid chemically-induced electron microscopy visible tag for marking specific proteins within the complex cellular environment. The pipeline provides efficient, distributable, low-cost sample preparation and enables targeted structural studies of identified proteins at the plasma membrane of mammalian cells.
External linksNat Commun / PubMed:39833141 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution7.5 - 19.8 Å
Structure data

EMDB-44909: Subtomogram average of 80S ribosome - consensus map
Method: EM (subtomogram averaging) / Resolution: 7.5 Å

EMDB-44921: Subtomogram average of 80S ribosome - non-rotated state
Method: EM (subtomogram averaging) / Resolution: 7.9 Å

EMDB-44922: Subtomogram average of 80S ribosome - rotated state
Method: EM (subtomogram averaging) / Resolution: 9.1 Å

EMDB-46973: Plasma membrane bound clathrin vertex (HEK293)
Method: EM (subtomogram averaging) / Resolution: 19.8 Å

Source
  • Homo sapiens (human)

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