关于人类D-3-糖酸盐脱酶的四级结构
Daniele Riva1, Marco Orlando1, Valentina Rabattoni1
1Department of Biotechnology and Life Sciences, University of Insubria, Varese, Italy.
Protein science : a publication of the Protein Society
|July 16, 2024
概括
D-3-糖酸脱酶 (PHGDH) 四聚体的形成对其稳定性和功能至关重要. ACT域中的突变破坏了这种组合,导致错误折叠和活动丧失,影响L-氨酸生物合成.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 酶学 是一种酶学.
背景情况:
- D-3-糖酸脱酶 (PHGDH) 对L-氨酸生物合成至关重要,这是与神经系统疾病相关的途径.
- 人类PHGDH (hPHGDH) 酶存在于同位四体,其C端具有调节性ACT和ASB域.
- 之前的结构研究仅限于hPHGDH的截断,二元形式.
研究的目的:
- 为了研究hPHGDH. tetrameric接口的特定残留的作用.
- 了解ACT和ASB领域对hPHGDH组装,稳定性和功能的贡献.
- 阐明hPHGDH正确折叠和活动的结构要求.
主要方法:
- 使用AlphaFold和分子动力学精细化进行计算建模,以预测四重体结构.
- 氨酸扫描突变发生,以研究选定的接口残留物的功能 (F418,R454,L478,P479,Y495).
- 对野生类型和突变变异的蛋白质活性,寡合状态,稳定性,折叠和聚合的分析.
主要成果:
- 创建了一个四度体hPHGDH模型,突出显示四度体界面的关键残留物.
- F418A变体 (ASB域) 对活性和四聚体形成产生了轻微影响,这表明它在二聚体识别中的作用.
- 在ACT域内的突变R454A,L478A,P479A和Y495A导致了四重体组件的丧失,错误折叠,聚合和活性降低.
结论:
- hPHGDH的四聚体接口,特别是ACT域中的残留物,对于正确的四聚体形成至关重要.
- 四化对于hPHGDH的正确折叠,稳定性和酶功能至关重要.
- 体组合的破坏对L-氨酸生物合成和细胞代谢有重大影响.
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