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单细胞蛋白循环动态的总体分析

Pierre Sabatier¹, Maico Lechner², Ulises H Guzmán²

¹Novo Nordisk Foundation Center for Protein Research, Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark; Cardio-Thoracic Translational Medicine (CTTM) Lab, Department of Surgical Sciences, Uppsala University, 752 37 Uppsala, Sweden.

|April 1, 2025

概括

单细胞蛋白质组学现在同时使用单细胞脉冲稳定同位素标记通过细胞培养中的氨基酸 (SC-pSILAC) 来测量蛋白质的丰度和循环. 这种方法揭示了细胞分化和蛋白质动态的新见解.

科学领域:

蛋白质组学
细胞生物学
生物化学

背景情况:

单细胞蛋白质组学 (SCP) 传统上侧重于蛋白质的丰富性,提供了有限的观点.
现有的SCP方法很大程度上是单维的,阻碍了细胞过程的全面分析.

研究的目的:

开发和验证一种新的单细胞蛋白质组学方法,用于同时测量蛋白质丰度和循环.
应用这种方法来研究人类诱导多能干细胞分化过程中的蛋白质动力学.

主要方法:

使用细胞培养中的氨基酸进行脉冲稳定同位素标记 (pSILAC) 进行单细胞分析 (SC-pSILAC).
应用最先进的SCP工作流程来分析单个HeLa细胞中的4000个蛋白质.
在两个月内进行了大规模的SC-pSILAC时间序列分析,对人类诱导多能干细胞 (iPSC) 进行了分化 (> 1,000 个细胞).

主要成果:

证明了检测两个SILAC标签的可行性,并在单细胞中量化约4000种蛋白质.
确定了蛋白质复合体的分化特异性联合调节和核心基因组的循环.
观察到基因组和一些细胞循环蛋白质不会随细胞直径变大,从而使细胞状态发生差异化.

结论:

通过SC-pSILAC方法,可以在单细胞水平上对蛋白质动态进行多维分析.
这种方法为细胞分化,蛋白质循环和细胞大小调节提供了新的见解.
通过提供更全面的蛋白质组视图,SC-pSILAC推动了单细胞生物学领域的发展.

关键词:

一个小小的埃沃塞普星际轨道飞行器一个细胞基因子 iPSC 的差异化质谱学蛋白质周转率脉冲的SILAC 单细胞蛋白质组学

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