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Updated: Jun 9, 2025

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线粒体蛋白质构成和相互作用的有针对性的分析,由内源性ROS触发的交叉连接器释放

Wen Zhou^1,2, Yuwan Chen^1,2, Wenxin Fu^1,3

¹State Key Laboratory of Medical Proteomics, National Chromatographic R. & A. Center, CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, 116023, China.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)

|October 30, 2024

概括

这项研究引入了针对线粒体的新型纳米粒子,以分析活细胞中的蛋白质构成和相互作用. 这种方法可以提高对线粒体蛋白质动态和生物功能的理解,而无需进行遗传修饰.

科学领域:

生物化学生物化学
细胞生物学细胞生物学
纳米技术纳米技术

背景情况:

线粒体蛋白质构成和相互作用研究对于理解细胞功能至关重要.
现有的化学交联质谱法 (CX-MS) 方法在分析未经修改的细胞中的线粒体蛋白质方面存在局限性.
在线粒体蛋白质动态的现场分析具有挑战性.

研究的目的:

开发一种用于现场分析线粒体蛋白质构成和相互作用的新方法.
克服当前CX-MS技术对线粒体蛋白质组学的局限性.
为了能够在没有遗传操纵的情况下对活细胞中的线粒体蛋白进行深入分析.

主要方法:

开发响应活性氧物种 (ROS) 的交叉链接器传递纳米粒子 (R-CDNP).
在R-CDNP中包含了线粒体向 (trifenylphosphine),ROS响应 (thioketal),加载 (PLGA) 和交叉链接器 (DSS) 模块.
在HepG2细胞中利用R-CDNP进行现场交叉链接和质谱分析.

主要成果:

在572个线粒体蛋白中成功识别了2103个交叉链接位点.
揭示了1718个内链,表明动态蛋白质构造,包括伴侣ATP依赖循环.
发现了385个相互联系,突出了OXPHOS复合体内的动态相互作用,并确定了27个潜在的新型相互作用.

结论:

R-CDNP使活细胞中线粒体蛋白质的动态构造和相互作用分析成为可能.
这种方法显著提高了对线粒体蛋白功能的理解.
R-CDNP平台为线粒体研究和疾病机制研究提供了一个强大的工具.

关键词:

交叉链接质谱法质谱法交叉链接质谱法线粒体中的线粒体.纳米颗粒是一种纳米粒子.蛋白质复合体蛋白质复合体反应性氧物种-响应性的反应性氧物种

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