Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA
Jing Cheng,
Tong Liu,
Xin You,
Fa Zhang,
Sen-Fang Sui,
Xiaohua Wan () and
Xinzheng Zhang ()
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Jing Cheng: Chinese Academy of Sciences
Tong Liu: Chinese Academy of Sciences
Xin You: Tsinghua University
Fa Zhang: Beijing Institute of Technology
Sen-Fang Sui: Tsinghua University
Xiaohua Wan: Beijing Institute of Technology
Xinzheng Zhang: Chinese Academy of Sciences
Nature Communications, 2023, vol. 14, issue 1, 1-9
Abstract:
Abstract Cryo-electron tomography is a major tool used to study the structure of protein complexes in situ. However, the throughput of tilt-series image data collection is still quite low. Here, we show that GisSPA, a GPU accelerated program, can translationally and rotationally localize the target protein complex in cellular lamellae, as prepared with a focused ion beam, using single cryo-electron microscopy images without tilt-series, and reconstruct the protein complex at near-atomic resolution. GisSPA allows high-throughput data collection without the acquisition of tilt-series images and reconstruction of the tomogram, which is essential for high-resolution reconstruction of asymmetric or low-symmetry protein complexes. We demonstrate the power of GisSPA with 3.4-Å and 3.9-Å resolutions of resolving phycobilisome and tetrameric photosystem II complex structures in cellular lamellae, respectively. In this work, we present GisSPA as a practical tool that facilitates high-resolution in situ protein structure determination.
Date: 2023
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DOI: 10.1038/s41467-023-36175-y
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