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开发和应用一个有针对性的光蛋白学方法来分析斑马鱼PAC2细胞系中的mTOR通路动态

Nikolai Huwa¹, René Schönenberger¹, Ksenia J Groh¹

¹Eawag, Swiss Federal Institute of Aquatic Science and Technology, 8600 Dübendorf, Switzerland.

Journal of proteomics

|June 4, 2025

概括

我们开发了一种抗体独立的方法来研究斑马鱼的mTOR途径,揭示了与哺乳动物相似的酸化动态. 这种方法使得比较生理学和毒理学领域的新研究成为可能.

科学领域:

细胞生物学细胞生物学
蛋白质组学是指蛋白质组学.
信号通道的信号通道

背景情况:

拉巴胺素 (mTOR) 途径的机械性标调节细胞生长,但由于抗体限制,其酸化动态在非哺乳动物模型中未得到充分研究.
斑马鱼 (Danio rerio) 是有价值的模型生物,但研究像mTOR这样的特定信号通路一直是具有挑战性的.

研究的目的:

开发和验证一种抗体独立的方法,用于量化斑马鱼的mTOR通路蛋白质酸化.
为了研究斑马鱼PAC2细胞在不同生长阶段的mTOR途径的酸化动态.

主要方法:

利用液体染色学-双重质谱法 (LC-MS/MS) 进行向的光蛋白质组学.
开发了优化的样本处理和数据分析策略,用于抗体独立量化.
斑马鱼PAC2细胞中的量化内源酸盐和蛋白质.

主要成果:

成功量化了与mTOR通路相关的10种内源酸盐和15种内源蛋白质.
观察到复杂的酸化动态,反映了细胞生长过渡 (指数式到静止式).
在斑马鱼和哺乳动物之间在mTOR通路结构和功能方面确认了整体相似性.

结论:

基于LC-MS/MS的向光蛋白学是一种可行且强大的方法,用于研究非哺乳动物物种的信号通路.
这种抗体独立的方法克服了各种生物体抗体可用性的局限性.
该方法支持环境监测的比较生理学,毒理学和生物标志物发现.

关键词:

这是LC-MS/MS.富含酸的丰富方法酸蛋白组学酸蛋白组学酸蛋白组学斑马鱼是一种斑马鱼.mTOR 信号传输系统

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