2023Cheng GisSPA

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Cheng, Jing / Liu, Tong / You, Xin / Zhang, Fa / Sui, Sen-Fang / Wan, Xiaohua / Zhang, Xinzheng. Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA. 2023. Nature Communications, Vol. 14, No. 1, p. 1282


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 highresolution in situ protein structure determination.




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