对MuRF1-UBE2复合物的结构预测确定了每个MuRF1-E2对相互作用选择性的氨基酸残留物
Agnès Claustre1, Mélodie Malige1, Maëlys Macheton1
1Université Clermont Auvergne, INRAE, UNH, Clermont-Ferrand, France.
The FEBS journal
|February 11, 2025
概括
肌肉缩E3联酶MuRF1与四个E2酶相互作用,通过计算和实验方法揭示了不同的接口. 了解这些相互作用可能会导致肌肉消耗疾病的新疗法.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- 在骨肌肉缩中,E3泛基因酶MuRF1 (TRIM63) 是至关重要的,它准收缩蛋白质.
- 在体外,MuRF1与四种不同的E2酶 (UBE2E1,UBE2J1,UBE2J2,UBE2L3) 相互作用,这表明它具有特殊的作用.
- 了解这些相互作用的分子基础是调节MuRF1活动的关键.
研究的目的:
- 研究MuRF1及其E2酶合作伙伴之间的分子接口.
- 阐明 MuRF1.1 通过选择性 E2 酶招募的结构基础.
- 根据MuRF1-E2相互作用来确定肌肉缩的潜在治疗点.
主要方法:
- 使用AlphaFold2和AlphaFold3进行计算建模,以预测复杂的接口.
- 表面等离子体共振 (SPR) 实验,以评估不同的灵敏度和结合亲和力.
- 使用点突变E2酶和截断的MuRF1.1进行亲和度测量.
- 基于光的测试检测相互作用诱导的变化.
主要成果:
- AlphaFold预测显示了每个MuRF1-E2复合体的独特接口残留物.
- SPR实验证实了明显的接口差异和结合特征.
- 亲和度测量验证了MuRF1-UBE2E1.1.的特定相互作用点,并确定了独特的相互作用.
- MuRF1-UBE2E1的相互作用可能涉及RING规范域外的域.
结论:
- MuRF1通过不同的接口表现出选择性的E2酶招募.
- MuRF1-UBE2E1相互作用具有独特的特征,可能涉及非RING域.
- 解读这些选择性相互作用为开发针对肌肉缩的向疗法提供了一条途径.
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