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-Structure paper
Title | Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA. |
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Journal, issue, pages | Nat Commun, Vol. 14, Issue 1, Page 1282, Year 2023 |
Publish date | Mar 15, 2023 |
![]() | Jing Cheng / Tong Liu / Xin You / Fa Zhang / Sen-Fang Sui / Xiaohua Wan / Xinzheng Zhang / ![]() |
PubMed 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 ...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. |
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Methods | EM (single particle) |
Resolution | 3.9 Å |
Structure data | ![]() EMDB-35976: Structure of in situ PSII from P. purpureum lamellae by GisSPA |
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