瑞士苏黎世联邦理工Evgeny Onischenko、Karsten Weis等研究人员合作利用代谢标记揭示多蛋白复合物的成熟动力学。该项研究成果于2020年12月16日在线发表在《细胞》杂志上。

研究人员提出了一种高通量策略来分析蛋白质复合物的天然组装动力学。研究人员应用这个方法来表征了320对构成酵母中≈50MDa核孔复合物（NPC）的核孔蛋白（NUP）的共装配。一些NUP通过快速交换来迅速组装，而另一些则需要漫长的成熟步骤。这揭示了NPC生物发生的分级原理，其中单个亚复合物在微小的时间尺度上形成，然后在大约1小时的成熟过程中从中心到外围共同组装。

有趣的是，NUP Mlp1的加入非常晚，并且优先与旧的NPC结合。这些方法在NPC之外很容易应用，从而有可能分析多种多蛋白装配体的细胞内动力学。

据介绍，所有蛋白质都与其他细胞成分相互作用来发挥其功能。尽管蛋白质复合物的鉴定已取得巨大进展，但其组装和动力学仍然难以表征。

附：英文原文

Title: Maturation Kinetics of a Multiprotein Complex Revealed by Metabolic Labeling

Author: Evgeny Onischenko, Elad Noor, Jonas S. Fischer, Ludovic Gillet, Matthias Wojtynek, Pascal Vallotton, Karsten Weis

Issue&Volume: 2020-12-16

Abstract: All proteins interact with other cellular components to fulfill their function. Whiletremendous progress has been made in the identification of protein complexes, theirassembly and dynamics remain difficult to characterize. Here, we present a high-throughputstrategy to analyze the native assembly kinetics of protein complexes. We apply ourapproach to characterize the co-assembly for 320 pairs of nucleoporins (NUPs) constitutingthe ≈50 MDa nuclear pore complex (NPC) in yeast. Some NUPs co-assemble fast via rapid exchangewhereas others require lengthy maturation steps. This reveals a hierarchical principleof NPC biogenesis where individual subcomplexes form on a minute timescale and thenco-assemble from center to periphery in a ～1 h-long maturation process. Intriguingly, the NUP Mlp1 stands out as joining verylate and associating preferentially with aged NPCs. Our approach is readily applicablebeyond the NPC, making it possible to analyze the intracellular dynamics of a varietyof multiprotein assemblies.

DOI: 10.1016/j.cell.2020.11.001

Source: https://www.cell.com/cell/fulltext/S0092-8674(20)31460-4