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在哺乳动物精子中使用现场冷电子断层扫描和AlphaFold2对接进行新的蛋白质鉴定

Zhen Chen¹, Momoko Shiozaki², Kelsey M Haas³

¹Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, USA; Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA.

|October 21, 2023

概括

这项研究引入了视觉蛋白质学工作流程,结合了冷电子断层扫描和AlphaFold2建模来揭示精子结构. 发现了新的蛋白质,Tektin 5

科学领域:

细胞生物学
结构生物学
蛋白质组学

背景情况:

了解细胞通路需要多种技术的方法来识别蛋白质,定位和确定结构.
哺乳动物的精子结构和功能依赖于轴突体内的复杂分子机制.

研究的目的:

开发和应用一个新的工作流程,将细胞冷电子断层扫描 (cryo-ET) 和AlphaFold2建模用于哺乳动物精子的现场结构分析.
识别与精子轴膜微管相关的新型蛋白质并阐明它们的功能.

主要方法:

用细胞冷电子断层扫描 (cryo-ET) 和亚断层扫描平均值来实现轴膜微管结构的高分辨率重建 (6.0 Å).
鼠标蛋白质组的AlphaFold2预测蛋白质模型与来自冷ET数据的精子特异密度进行了计算匹配.
进行了Tektin 5 (Tekt5-/ -) 的基因淘汰,以调查其体内功能.

主要成果:

高分辨率的冷ET重建通过与AlphaFold2模型匹配,使精子特异性蛋白质的无偏见识别成为可能.
鉴定出Tektin 5,CCDC105和SPACA9是精子轴突中的新型微管相关蛋白质.
这些蛋白质形成一个交互网络,交叉连接双管微管,影响它们的机械特性,正如Tekt5-/-精子中变形的鞭毛所证明的那样.

结论:

这项研究为现场结构生物学提供了强大的细胞视觉蛋白质学工作流程.
鉴定到的蛋白质,特别是Tektin 5, 在保持精子鞭状结构和机械完整性方面起着至关重要的作用.
这项研究有助于我们更好地了解哺乳动物精子成分的分子结构和体内功能.

关键词:

阿尔法Fold2建模一个字母细胞冷ET 双人车微管子微管内蛋白质精子视觉蛋白质组学

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