Cryo-electron tomography pipeline for plasma membranes

Willy W. Sun, Dennis J. Michalak, Kem A. Sochacki (), Prasanthi Kunamaneni, Marco A. Alfonzo-Méndez, Andreas M. Arnold, Marie-Paule Strub, Jenny E. Hinshaw () and Justin W. Taraska ()
Additional contact information
Willy W. Sun: US National Institutes of Health
Dennis J. Michalak: US National Institutes of Health
Kem A. Sochacki: US National Institutes of Health
Prasanthi Kunamaneni: US National Institutes of Health
Marco A. Alfonzo-Méndez: US National Institutes of Health
Andreas M. Arnold: US National Institutes of Health
Marie-Paule Strub: US National Institutes of Health
Jenny E. Hinshaw: US National Institutes of Health
Justin W. Taraska: US National Institutes of Health

Nature Communications, 2025, vol. 16, issue 1, 1-14

Abstract: Abstract 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.

Date: 2025
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DOI: 10.1038/s41467-025-56045-z

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