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Smart Molecular Nanosheets for Advanced Preparation of Biological Samples in Electron Cryo-Microscopy.
ACS Nano ( IF 15.8 ) Pub Date : 2020-06-26 , DOI: 10.1021/acsnano.0c03052 Julian Scherr 1 , Zian Tang 2 , Maria Küllmer 2 , Sebastian Balser 1 , Alexander Stefan Scholz 1 , Andreas Winter 2 , Kristian Parey 3 , Alexander Rittner 4 , Martin Grininger 4 , Volker Zickermann 5 , Daniel Rhinow 3 , Andreas Terfort 1 , Andrey Turchanin 2
ACS Nano ( IF 15.8 ) Pub Date : 2020-06-26 , DOI: 10.1021/acsnano.0c03052 Julian Scherr 1 , Zian Tang 2 , Maria Küllmer 2 , Sebastian Balser 1 , Alexander Stefan Scholz 1 , Andreas Winter 2 , Kristian Parey 3 , Alexander Rittner 4 , Martin Grininger 4 , Volker Zickermann 5 , Daniel Rhinow 3 , Andreas Terfort 1 , Andrey Turchanin 2
Affiliation
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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.
中文翻译：
智能分子纳米片，用于电子低温显微镜中生物样品的高级制备。
陶瓷化的生物标本的透射电子低温显微镜（cryoEM）是结构生物学的强大工具。目前，玻璃化生物样品的制备始于样品分离和纯化，然后固定在独立的无定形冰层中。在这里，我们证明了具有特定生物识别位点的超薄（〜10 nm）智能分子纳米片嵌入到生物排斥层中，该生物排斥层与机械稳定的碳纳米膜共价结合，从而使分离和结构分析变得更加简单。我们使用X射线光电子和红外光谱，原子力显微镜以及表面等离振子共振测量等互补方法，详细表征了这些纳米片的工程及其生物识别特性。从混合物中原位选择带有His标签的蛋白质，然后通过cryoEM分析其结构。
更新日期：2020-08-25
中文翻译：
智能分子纳米片，用于电子低温显微镜中生物样品的高级制备。
陶瓷化的生物标本的透射电子低温显微镜（cryoEM）是结构生物学的强大工具。目前，玻璃化生物样品的制备始于样品分离和纯化，然后固定在独立的无定形冰层中。在这里，我们证明了具有特定生物识别位点的超薄（〜10 nm）智能分子纳米片嵌入到生物排斥层中，该生物排斥层与机械稳定的碳纳米膜共价结合，从而使分离和结构分析变得更加简单。我们使用X射线光电子和红外光谱，原子力显微镜以及表面等离振子共振测量等互补方法，详细表征了这些纳米片的工程及其生物识别特性。从混合物中原位选择带有His标签的蛋白质，然后通过cryoEM分析其结构。
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