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通过时间解决的过渡金属离子FRET揭示了Mfn1中GTP合的构造变化的结构洞察力

S M Hurwitz¹, W N Zagotta², S E Gordon²

¹Department of Biochemistry, University of Washington, Seattle, WA 98195.

bioRxiv : the preprint server for biology

|November 24, 2025

概括

使用时间解析的FRET研究了米托素1 (Mfn1) 动态. 对Mfn1的GTP结合有利于开放状态,揭示了对线粒体融合机制的新见解.

科学领域:

生物化学生物化学
分子生物学分子生物学
细胞生物学细胞生物学

背景情况:

线粒体外膜融合依赖于线粒体素 (Mfn1和Mfn2).
了解Mfn1的核酸驱动的结构变化对于阐明膜融合机制至关重要.
目前的结构数据是不完整的,留下了的机制神秘.

研究的目的:

为了研究Mfn1.1.的GTP合的结构动态.
描述Mfn1在整个催化循环中的结构状态.
为Mfn1的GTP驱动的形状变化的结构和能量提供基本的见解.

主要方法:

利用时间解析的过渡金属离子光共振能量转移 (tmFRET).
在最小的Mfn1构造 (GTPase域和HB1通过Hinge2连接) 上设计的FRET对.
用光寿命测量tmFRET,以确定不同核酸结合状态的距离分布.

主要成果:

在溶液中确认了与GDP相关的Mfn1的开放状态.
揭示了过渡状态是在GDP+Pi的存在下,开放和封闭状态之间的平衡.
首次捕获了Mfn1的GTP结合和无核酸状态.
证明GTP结合有利于开放状态,表明一种新的结构变化.

结论:

由于GTP结合和水解,Mfn1经历了显著的结构变化.
Mfn1的apo,GDP-bound和GTP-bound状态表现出不同的形状.
这些发现为Mfn1在线粒体融合中的全性机制提供了基本的见解.

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