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Updated: Jan 8, 2026

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An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity
Published on: October 8, 2018
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通过非典型的SPOUT甲基转移酶Trm10进行tRNA基质识别和修饰的分子基础
Suparno Nandi1, Sarah E Strassler1, Debayan Dey1
1Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA.
bioRxiv : the preprint server for biology
|December 22, 2025
概括
Trm10甲基转移酶使用特定的相互作用来修改tRNA中的guanosin 9. 化EM结构揭示了Trm10如何结合tRNA,引导瓜诺辛进行甲基化,并显示了瓜诺辛对腺的选择性.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 分子生物学分子生物学
背景情况:
- Trm10是一种进化保守的甲基转移酶.
- 它在特定转移RNA (tRNA) 中修改了瓜诺辛9 (G9) 的N1位置.
- 对于Trm10基质的识别和修饰的机制尚不清楚.
研究的目的:
- 为了阐明Trm10介导的tRNA修饰的结构基础.
- 了解酶的基质识别和瓜诺辛选择性.
- 为了揭示一个RNA修饰酶Trm10的作用机制.
主要方法:
- 低温电子显微镜 (cryo-EM) 用于确定Trm10-tRNA复合物的结构.
- 使用一种S-adenosyl-L-methionine (SAM) 类比物来捕获催化后复合物.
- 进行了分子动力学模拟,以比较G9和A9结合.
主要成果:
- 观察到三种不同的Trm10-tRNA复合体:单体 ("封闭"和"开放"构造) 和二极体.
- 单体复合物表现出具有正电荷的表面和稳定G9.9的"状"相互作用.
- 分子动力学模拟揭示了瓜诺辛选择性的结构基础,G9在A9.9上稳定.
结论:
- 这项研究揭示了G9特异性tRNA通过Trm10.0甲基化的结构决定因素.
- 在SPOUT甲基转移酶中提出了Trm10的独特作用机制.
- 这些发现提供了对tRNA-蛋白相互作用和酶基质识别的见解.
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