Rose Kevin, Herrmann Eric, Kakudji Eve, Lizarrondo Javier, Celebi A Yasemin, Wilfling Florian, Lewis Samantha C, Hurley James H
Aligning Science Across Parkinson's Collaborative Research Network, Chevy Chase, MD 20815.
California Institute for Quantitative Biosciences, University of California Berkeley, Berkeley, CA 94720.
Proc Natl Acad Sci U S A. 2025 Aug 5;122(31):e2511890122. doi: 10.1073/pnas.2511890122. Epub 2025 Jul 31.
Defective mitochondrial quality control in response to loss of mitochondrial membrane polarization is implicated in Parkinson's disease by mutations in and . Parkin-expressing U2 osteosarcoma (U2OS) cells were treated with the depolarizing agents oligomycin and antimycin A (OA) and subjected to cryo-focused ion beam milling and in situ cryo-electron tomography. Mitochondria were fragmented and devoid of matrix calcium phosphate crystals. Phagophores were visualized, with bridge-like lipid transporter densities connected to mitophagic phagophores. A subpopulation of ATP synthases relocalized from cristae to the inner boundary membrane. The structure of the dome-shaped prohibitin complex, a dodecamer of PHB1-PHB2 dimers, was determined in situ by subtomogram averaging in untreated and treated cells and found to exist in open and closed conformations, with the closed conformation being enriched by OA treatment. These findings provide a set of native snapshots of the manifold nano-structural consequences of mitochondrial depolarization and provide a baseline for future in situ dissection of Parkin-dependent mitophagy.
线粒体膜电位丧失时线粒体质量控制缺陷与帕金森病相关，这与 和 中的突变有关。用去极化剂寡霉素和抗霉素A（OA）处理表达帕金蛋白的U2骨肉瘤（U2OS）细胞，并进行低温聚焦离子束铣削和原位低温电子断层扫描。线粒体发生碎片化，且没有基质磷酸钙晶体。观察到吞噬泡，有与线粒体自噬吞噬泡相连的桥状脂质转运体密度。一部分ATP合酶从嵴重新定位到内膜边界膜。通过对未处理和处理细胞的亚断层平均原位确定了穹顶状抑制素复合物（一种由PHB1 - PHB2二聚体组成的十二聚体）的结构，发现其以开放和闭合构象存在，OA处理后闭合构象增多。这些发现提供了一组线粒体去极化多种纳米结构后果的天然快照，并为未来原位剖析帕金蛋白依赖性线粒体自噬提供了基线。
