ShanghaiTech University Knowledge Management System
| VitisC: Visualizing interested target via integrated scaffold of cytoophidium | |
| 2025-01-19 | |
| 状态 | 已发表 |
| 摘要 | Cryo-electron microscopy (cryo-EM) single-particle analysis has become a widely used technique for high-resolution structural determination of biological macromolecules and complexes. However, the determination of the structure of small molecule proteins remains a limitation of this technology. To address this issue, here we develop a novel approach termed Visualizing interested target via integrated scaffold of cytoophidium (VitisC). We use a filamentous structure formed by Escherichia coli CTP synthase (CTPS) as the scaffold, termed the scaffold of cytoophidium. Through artificial design and modification, small proteins can be attached to this symmetrical scaffold, which is very suitable for cryo-EM imaging. The formation conditions for stable filament structures of the fusion protein are optimized in vitro, and the three-dimensional structure of the fusion protein is reconstructed using cryo-EM single-particle analysis, achieving an overall resolution of 3.39 Ã . Therefore, cryo-EM is successfully applied to visualize small proteins. VitisC not only demonstrates the feasibility of employing cytoophidia as scaffolds, but also provides an approach for high-resolution structural analysis of small proteins using Cryo-EM. |
| 关键词 | cryo-EM CTP synthase cytoophidium the scaffold of cytoophidium VitisC |
| 语种 | 英语 |
| DOI | 10.1101/2025.01.17.633574 |
| 相关网址 | 查看原文 |
| 出处 | bioRxiv |
| 收录类别 | PPRN.PPRN |
| WOS记录号 | PPRN:120823936 |
| WOS类目 | Biochemistry & Molecular Biology |
| 资助项目 | National Natural Science Foundation of China[32350710195] ; UK Medical Research Council["MC_UU_12021/3","474 MC_U137788471"] |
| 文献类型 | 预印本 |
| 条目标识符 | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/503597 |
| 专题 | 生命科学与技术学院 生命科学与技术学院_PI研究组_刘冀珑组 生命科学与技术学院_博士生 |
| 通讯作者 | Liu, Ji-Long |
| 作者单位 | 1.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China 2.Univ Oxford, Dept Physiol Anat & Genet, Oxford OX19 3PT, England |
| 推荐引用方式 GB/T 7714 | Yin, Boqi,Hu, Huan-Huan,Zhong, Jiale,et al. VitisC: Visualizing interested target via integrated scaffold of cytoophidium. 2025. |
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