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使用15-YNE的细胞标记对于在同一样本中跟踪蛋白质棕化和代谢脂质流量是有用的

Nadine Merz¹, Karin Schilling¹, Dominique Thomas^1,2,3

¹Goethe University Frankfurt, Institute of Clinical Pharmacology, Faculty of Medicine, Theodor Stern Kai 7, 60590 Frankfurt am Main, Germany.

Molecules (Basel, Switzerland)

|January 25, 2025

概括

用15-hexadecynoic acid (15-YNE) 的代谢标签有效地跟踪蛋白质S-palmitoylation和脂质代谢. 这种可追溯的脂质方法有助于同时研究这两种过程,而不会改变细胞脂质组成.

科学领域:

生物化学生物化学
细胞生物学细胞生物学
蛋白质组学是指蛋白质组学.

背景情况:

蛋白质S-palmitoylation涉及通过一个 thioester 键将脂肪酸连接到cysteine.
有各种方法可以检测蛋白质S-palmitoylation.
可追溯脂质标签对细胞脂质代谢和脂质组的影响需要研究.

研究的目的:

用15-hexadecynoic acid (15-YNE) 进行代谢标记后检测蛋白S-palmitoylation的两种方法进行比较.
开发一种方法来追踪15-YNE在同一样本中的脂质和蛋白质的结合.
评估15-YNE标签对细胞脂质代谢和脂质组的影响.

主要方法:

用15-hexadecynoic acid (15-YNE) 对细胞进行代谢标记.
液体染色学高分辨率质谱学 (LC-HRMS) 用于追踪15-YNE在脂质中.
使用可点击的化物或生物酸盐检测蛋白质S-palmitoylation,然后进行免疫沉.

主要成果:

观察到时间和度依赖的calnexin和SDHA的S-palmitoylation.
可点击的化物和生物酸方法都被证明对S-palmitoylation检测同样有用.
15-YNE被纳入了各种脂类,但没有显著改变整体脂质组成.

结论:

15-YNE是同时检测蛋白S-palmitoylation和脂质代谢的有价值的标记物.
开发的LC-HRMS方法允许同时分析脂质合并和蛋白质棕化.
使用15-YNE的可追溯脂质标签为研究脂质-蛋白质相互作用提供了强大的方法.

关键词:

亚酸 Cy5.5-化物这是LC-HRMS.在Linex上,Linex是最重要的.基因是一种基因.生物生物是一种生物.点击反应点击反应反应蛋白质棕化转化为棕化

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