Nature Communications, 15 March, 2023, DOI:https://doi.org/10.1038/s41467-023-36175-y
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 & Xinzheng Zhang
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.
文章链接:https://www.nature.com/articles/s41467-023-36175-y
相关报道:http://www.ibp.cas.cn/kyjz/zxdt/202303/t20230316_6698172.html
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