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Smart Molecular Nanosheets for Advanced Preparation of Biological Samples in Electron Cryo-Microscopy

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Parey,  Kristian
Department of Structural Biology, Max Planck Institute of Biophysics, Max Planck Society;

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Rhinow,  Daniel
Department of Structural Biology, Max Planck Institute of Biophysics, Max Planck Society;

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Citation

Scherr, J., Tang, Z., Küllmer, M., Balser, S., Scholz, A. S., Winter, A., et al. (2020). Smart Molecular Nanosheets for Advanced Preparation of Biological Samples in Electron Cryo-Microscopy. ACS Nano, 14(8), 9972-9978. doi:10.1021/acsnano.0c03052.


Cite as: http://hdl.handle.net/21.11116/0000-0006-AB7C-F
Abstract
Transmission electron cryo-microscopy (cryoEM) of vitrified biological specimens is a powerful tool for structural biology. Current preparation of vitrified biological samples starts off with sample isolation and purification, followed by the fixation in a freestanding layer of amorphous ice. Here, we demonstrate that ultrathin (∼10 nm) smart molecular nanosheets having specific biorecognition sites embedded in a biorepulsive layer covalently bound to a mechanically stable carbon nanomembrane allow for a much simpler isolation and structural analysis. We characterize in detail the engineering of these nanosheets and their biorecognition properties employing complementary methods such as X-ray photoelectron and infrared spectroscopy, atomic force microscopy as well as surface plasmon resonance measurements. The desired functionality of the developed nanosheets is demonstrated by in situ selection of a His-tagged protein from a mixture and its subsequent structural analysis by cryoEM.