基于TurboID的近距离标签揭示了VPS34和细胞平衡之间存在联系
Taotao Wang1, Xinjing Li1, Ningjing Liu1
1State Key Laboratory of Microbial Metabolism & Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, PR China.
Journal of plant physiology
|September 25, 2023
概括
植物和酵母在细胞贩运中使用III类酸丁3-激酶 (PI3KC3). 这项研究揭示了VPS34,PI3KC3子单元,与碳水化合物代谢和膜潜能蛋白相互作用,将PI3KC3与细胞平衡联系起来.
科学领域:
- 细胞生物学 细胞生物学
- 植物生物化学 植物生物化学
背景情况:
- 植物和酵母有一种单一的III类酸丁3-激酶 (PI3KC3),与动物不同.
- PI3KC3的脂质产物,酸-3酸 (PI3P),对于细胞内贩运途径至关重要,包括自细胞和多细胞体的形成.
- 植物PI3KC3的催化子单元是VPS34,对细胞平衡和发育至关重要,但它的相互作用体仍然没有特征.
研究的目的:
- 通过近距离标记来识别Arabidopsis VPS34的近距离互动体.
- 为了揭示VPS34的新功能和相互作用,超出已知的内溶酶体贩运作用.
- 研究PI3KC3与细胞过程 (如碳水化合物代谢和膜潜力) 之间的联系.
主要方法:
- 利用TurboID,一种酶催化近距离标记方法,用于空间限制的生物化.
- 使用亲和力净化与质谱相结合,以丰富和识别与VPS34相互作用的蛋白质.
- 在植物中使用生物化学和细胞分析验证了特定蛋白质相互作用.
主要成果:
- 产生了阿拉比多普西斯VPS34的近端互动组,识别了273种相关蛋白质.
- 证实了VPS34在内溶酶体贩运途径中的已知作用.
- 发现了与碳水化合物代谢相关的蛋白质的显著丰富,包括甘油酶 (例如GAPDH) 和糖糖生物合成酶.
- 验证了VPS34与甘甲基-3-酸脱酶 (GAPDH) 和血膜H+-ATPase AHA2.2之间的相互作用.
结论:
- 这项研究提供了VPS34的第一个全面的互动组,扩大了我们对植物PI3KC3功能的理解.
- 通过VPS34将PI3KC3与关键的细胞过程联系起来,包括碳水化合物代谢和膜潜力的调节.
- 突出了PI3KC3在通过这些新发现的相互作用来维持细胞平衡中的新角色.
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