ジャーナル: Nat Commun / 年: 2025 タイトル: Cryo-electron tomography pipeline for plasma membranes. 著者: Willy 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 / 要旨: 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 ...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.