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Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA

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 c...

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Autores principales: Cheng, Jing, Liu, Tong, You, Xin, Zhang, Fa, Sui, Sen-Fang, Wan, Xiaohua, Zhang, Xinzheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10017804/
https://www.ncbi.nlm.nih.gov/pubmed/36922493
http://dx.doi.org/10.1038/s41467-023-36175-y
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author Cheng, Jing
Liu, Tong
You, Xin
Zhang, Fa
Sui, Sen-Fang
Wan, Xiaohua
Zhang, Xinzheng
author_facet Cheng, Jing
Liu, Tong
You, Xin
Zhang, Fa
Sui, Sen-Fang
Wan, Xiaohua
Zhang, Xinzheng
author_sort Cheng, Jing
collection PubMed
description 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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spelling pubmed-100178042023-03-17 Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA Cheng, Jing Liu, Tong You, Xin Zhang, Fa Sui, Sen-Fang Wan, Xiaohua Zhang, Xinzheng Nat Commun Article 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. Nature Publishing Group UK 2023-03-15 /pmc/articles/PMC10017804/ /pubmed/36922493 http://dx.doi.org/10.1038/s41467-023-36175-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
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
title Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA
title_full Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA
title_fullStr Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA
title_full_unstemmed Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA
title_short Determining protein structures in cellular lamella at pseudo-atomic resolution by GisSPA
title_sort determining protein structures in cellular lamella at pseudo-atomic resolution by gisspa
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10017804/
https://www.ncbi.nlm.nih.gov/pubmed/36922493
http://dx.doi.org/10.1038/s41467-023-36175-y
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