AlphaFold2有助于提供新的机械洞察力,了解LUBAC子单元之间的相互作用
Chenchen Wang1, Chunying Gu2, Ying Lv3
1College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
Acta biochimica et biophysica Sinica
|April 24, 2024
概括
卢巴克 (LUBAC) 综合体是一个复杂的建筑.
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:
- 线性ubiquitin链组合复合体 (LUBAC) 对于细胞信号传输至关重要,并且是唯一具有特定子单元相互作用的E3酶复合体.
- 众所周知,SHARPIN和HOIL-1L的泛素类 (UBL) 域与HOIP相互作用,从而有助于LUBAC的结构完整性.
- 在介导SHARPIN/HOIL-1L UBL域与HOIP之间的相互作用中,LTM图案的确切作用仍然不完全理解.
研究的目的:
- 研究LTM图案对LUBAC复合体内的结构稳定性和子单元相互作用的影响.
- 为了阐明HOIP,HOIL-1L-UBL和HOIP (466695) 结构之间的结合机制.
主要方法:
- 利用AlphaFold2来预测带有和没有LTM动机的LUBAC结构.
- 使用 GeoPPI 来计算从单点突变中结合的自由能量变化 (ΔG),模拟蛋白质-蛋白质相互作用.
- 进行了Gromacs分子动力学模拟,表面等离子体共振 (SPR) 和异热定位热量测量 (ITC) 实验.
主要成果:
- 在 AlphaFold2 预测的结构中缺少 LTM 图案会改变 HOIP-UBA 结构.
- 这种LTM图案降低了SHARPIN/HOIL-1L UBL域与HOIP之间的相互作用,降低了LUBAC的结构稳定性.
- HOIP (629695) 和HOIP-UBA与HOIL-1L-UBL结合,形成一个稳定的复合体 (HOIP (466695) 结构).
结论:
- LTM 图案负面调节了 UBL 域与 HOIP 之间的相互作用,影响了 LUBAC 的结构稳定性.
- 对于LUBAC的结构完整性来说,HOIP-UBA,HOIP (629695) 和HOIL-1L-UBL之间的协作互动是必不可少的.
- 结果为LUBAC复杂组装和调节提供了新的见解.
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