STAgSPA策略原位测定蓝藻藻胆体-PSII超复合物的结构-动脉网

In situ structural determination of cyanobacterial phycobilisome–PSII supercomplex by STAgSPA strategy

Nature 等信源发布 2024-08-22 11:50

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AbstractPhotosynthesis converting solar energy to chemical energy is one of the most important chemical reactions on earth. In cyanobacteria, light energy is captured by antenna system phycobilisomes (PBSs) and transferred to photosynthetic reaction centers of photosystem II (PSII) and photosystem I (PSI).

光合作用将太阳能转化为化学能是地球上最重要的化学反应之一。在蓝藻中，光能被天线系统藻胆体（PBS）捕获，并转移到光系统II（PSII）和光系统I（PSI）的光合反应中心。

While most of the protein complexes involved in photosynthesis have been characterized by in vitro structural analyses, how these protein complexes function together in vivo is not well understood. Here we implemented STAgSPA, an in situ structural analysis strategy, to solve the native structure of PBS–PSII supercomplex from the cyanobacteria Arthrospira sp.

。在这里，我们实施了原位结构分析策略STAgSPA，以解决蓝藻Arthrospira sp。中PBS–PSII超复合物的天然结构。

FACHB439 at resolution of ~3.5 Å. The structure reveals coupling details among adjacent PBSs and PSII dimers, and the collaborative energy transfer mechanism mediated by multiple super-PBS in cyanobacteria. Our results provide insights into the diversity of photosynthesis-related systems between prokaryotic cyanobacteria and eukaryotic red algae but are also a methodological demonstration for high-resolution structural analysis in cellular or tissue samples..

FACHB439的分辨率约为3.5Å。该结构揭示了相邻PBS和PSII二聚体之间的耦合细节，以及蓝藻中多个超级PBS介导的协同能量转移机制。我们的结果提供了对原核蓝藻和真核红藻之间光合作用相关系统多样性的见解，但也是细胞或组织样品中高分辨率结构分析的方法学证明。。

IntroductionIn oxygenic photosynthetic organisms, the light-driven reactions depend on the cooperation of different multiprotein complexes that are attached to or embedded in thylakoid membranes, such as antenna systems, photosystems II/I (PSII/PSI) and cytochrome b6f (Cyt b6f)1,2. In cyanobacteria, sunlight is harvested by the soluble light-harvesting apparatus, phycobilisomes (PBSs), and the energy is finally transferred to the reaction centers of PSII/PSI to induce photo-induced electron transport3,4.

引言在产氧光合生物中，光驱动反应取决于附着或嵌入类囊体膜的不同多蛋白复合物的协同作用，如天线系统，光系统II/I（PSII/PSI）和细胞色素b6f（Cyt b6f）1,2。在蓝藻中，阳光通过可溶性光收集装置藻胆体（PBS）收集，能量最终转移到PSII/PSI的反应中心以诱导光诱导电子传输3,4。

The structures of isolated PBS, PSII, and PSI have been determined individually by cryo-electron microscopy (cryo-EM) single-particle analysis (SPA) at near-atomic resolution5,6,7,8, providing the structural basis for energy transfer, electron transfer, and photoprotection within the isolated complexes.

分离的PBS，PSII和PSI的结构已经通过低温电子显微镜（cryo-EM）单粒子分析（SPA）在近原子分辨率下分别测定5,6,7,8，为能量转移，电子转移和光保护提供了结构基础。在分离的复合物中。

However, it is hard to obtain intact assemblies of these protein machinery by SPA due to their diverse protein properties, which may result in the dissociation of some loosely associated supercomplexes during purification. Previously, the cross-linking coupled with mass spectrometry analysis has identified some potential interaction sites of PBS–PSII–PSI megacomplex9.

然而，由于它们不同的蛋白质特性，很难通过SPA获得这些蛋白质机器的完整组装体，这可能导致纯化过程中一些松散相关的超复合物的解离。以前，交联结合质谱分析已经确定了PBS–PSII–PSI巨复合物9的一些潜在相互作用位点。

The low-resolution negative-staining PBS–PSII and PBS–CpcL–PSI structures also indicate the coarse binding patterns between PBS and PSII/PSI in cyanobacteria10,11. However, the exact interactions and energy transfer pathways of PBS–PSII/PSI at their native state remain elusive.Cryo-electron tomography (Cryo-ET) coupled with subtomogram averaging (STA) is emerging as a potent methodology for addressing target particles in the cellular environment, which has been applied to visualize the arrangement of PBSs, PSII, or ATPase in thylakoid membranes12,13.

。但是，PBS–PSII/PSI在其天然状态下的确切相互作用和能量转移途径仍然难以捉摸。冷冻电子断层扫描（Cryo-ET）与亚谱平均（STA）相结合，正在成为一种有效的方法，用于解决细胞环境中的目标颗粒，已被用于可视化类囊体膜中PBS，PSII或ATPase的排列12,13。

Although some complexes, i.e., ribosomes14, cilia15, and .

尽管有些复合物，即核糖体14，纤毛15和。

Data availability

数据可用性

The cryo-EM density maps and atomic models have been deposited in the Electron Microscopy Data Bank and the Protein Data Bank under the following accession codes: 8WQL (PBS–PSII supercomplex structure), EMD-37749 (artificially stitched map), EMD-37738 (local map of PBS at 3.5 Å resolution), EMD-37747 (local map of PSII dimer at 3.6 Å resolution), and EMD-37750 (low-resolution map of PBS–PSII at 5.1 Å resolution).

低温电磁密度图和原子模型已以以下登录号保存在电子显微镜数据库和蛋白质数据库中：8WQL（PBS-PSII超复合物结构），EMD-37749（人工缝合图），EMD-37738（PBS的局部图，分辨率为3.5Å），EMD-37747（PSII二聚体的局部图，分辨率为3.6Å）和EMD-37750（PBS-PSII的低分辨率图，分辨率为5.1Å）。

References structures used in this work are 7RCV, 7EXT, and EMD-4602. Source data are provided in this paper..

这项工作中使用的参考结构是7RCV，7EXT和EMD-4602。本文提供了源数据。。

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Download referencesAcknowledgementsWe thank the staff at the Tsinghua University Branch of the National Protein Science Facility (Beijing) for their technical support on the cryo-EM and high-performance computation platforms. We thank Dr. Hong-Wei Wang for providing inspiring discussion for the cryo-EM study in this work.

下载参考文献致谢我们感谢国家蛋白质科学基金（北京）清华大学分校的工作人员在低温电磁和高性能计算平台上提供的技术支持。我们感谢王洪伟博士为这项工作中的低温电磁研究提供了鼓舞人心的讨论。

We thank Dr. Wei Lu from the Department of Chemistry, Southern University of Science and Technology, for his assistance on 77 K fluorescence spectra. This work was supported by the National Natural Science Foundation of China (32241030 and 32271245 to S.-F.S.), the Young Elite Scientist Sponsorship Program by CAST (2023QNRC001 to X.Y.), China Postdoctoral Science Foundation (2023M741985 to X.Y.), and the National Basic Research Program (2016YFA0501101 and 2017YFA0504600 to S.-F.S.).Author informationAuthor notesThese authors contributed equally: Xing Zhang, Yanan Xiao, Xin You.Authors and AffiliationsMinistry of Education Key Laboratory of Protein Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Frontier Research Center for Biological Structures, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing, ChinaXing ZhangSchool of Life Sciences, Southern University of Science and Technology, Shenzhen, ChinaYanan Xiao & Sen-Fang SuiState Key Laboratory of Membrane Biology, Beijing Frontier Research Center for Biological Structures, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing, ChinaXin You, Shan Sun & Sen-Fang SuiAuthorsXing ZhangView author publicationsYou can also search for this author in.

我们感谢南方科技大学化学系的Wei Lu博士在77 K荧光光谱方面的帮助。这项工作得到了国家自然科学基金（3224100和32271245至S.-F.S.），CAST青年精英科学家赞助计划（2023QNRC001至X.Y.），中国博士后科学基金（2023M741985至X.Y.）和国家基础研究计划（2016YFA050101和2017YFA050400至S.-F.S.）的支持。作者信息作者注意到这些作者做出了同样的贡献：张兴，肖亚南，辛友。作者和所属单位清华大学生命科学学院蛋白质科学教育重点实验室，清华-北京生命科学联合中心，北京生物结构前沿研究中心，北京结构生物学高级创新中心，清华大学生命科学学院，北京，张兴南方科技大学生命科学学院，深圳，肖亚南和隋森芳膜生物学国家重点实验室，北京生物结构前沿研究中心，北京结构生物学高级创新中心，清华大学生命科学学院，北京，中国信友，孙珊芳和隋森芳作者张兴观点作者出版物您也可以在中搜索这位作者。

PubMed Google ScholarYanan XiaoView author publicationsYou can also search for this author in

PubMed Google ScholarYanan XiaoView作者出版物您也可以在

PubMed Google ScholarXin YouView author publicationsYou can also search for this author in

PubMed Google ScholarXin YouView作者出版物您也可以在

PubMed Google ScholarShan SunView author publicationsYou can also search for this author in

PubMed Google ScholarShan SunView作者出版物您也可以在

PubMed Google ScholarSen-Fang SuiView author publicationsYou can also search for this author in

PubMed Google ScholarSen Fang SuiView作者出版物您也可以在

PubMed Google ScholarContributionsS.-F.S. supervised the project. Y.X. and X.Y. froze samples, and performed cryo-FIB milling and sequence analysis. X.Z. designed the cryo-EM strategy and optimized the data collection scripts. X.Z., X.Y., and Y.X. collected the cryo-EM data. X.Z.

PubMed谷歌学术贡献-F、美国监督了该项目。Y、 X.和X.Y.冻结样品，并进行cryo-FIB铣削和序列分析。十、 Z.设计了cryo-EM策略并优化了数据收集脚本。十、 Z.，X.Y。和Y.X.收集了低温电磁数据。十、 Z。

performed the cryo-EM analysis. X.Y. and Y.X. performed the model building and the structure refinement. Y.X. performed the biochemical experiments. X.Y., Y.X., X.Z., S.S., and S.-F.S. analyzed the structure; all authors contributed in writing the manuscript.Corresponding authorsCorrespondence to.

进行了低温电磁分析。十、 Y.和Y.X.进行了模型构建和结构改进。Y、 X.进行了生化实验。十、 Y.，Y.X.，X.Z.，S.S。和S.-F.S.分析了结构；所有作者都为撰写手稿做出了贡献。通讯作者通讯。

Xing Zhang or Sen-Fang Sui.Ethics declarations

张兴或隋森芳。道德宣言

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Reprints and permissionsAbout this articleCite this articleZhang, X., Xiao, Y., You, X. et al. In situ structural determination of cyanobacterial phycobilisome–PSII supercomplex by STAgSPA strategy.

转载和许可本文引用本文Zhang，X.，Xiao，Y.，You，X。等人。通过STAgSPA策略原位测定蓝藻藻胆体-PSII超复合物的结构。

Nat Commun 15, 7201 (2024). https://doi.org/10.1038/s41467-024-51460-0Download citationReceived: 29 November 2023Accepted: 08 August 2024Published: 22 August 2024DOI: https://doi.org/10.1038/s41467-024-51460-0Share this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

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生物能量冷冻电子断层摄影合成

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