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通过代谢标记揭示多蛋白复合体的成熟动力学

Evgeny Onischenko¹, Elad Noor², Jonas S Fischer³

¹Department of Biological Sciences, University of Bergen, Bergen 5020, Norway; Institute of Biochemistry, Department of Biology, ETH Zurich, Zurich CH-8093, Switzerland.

|December 17, 2020

概括

研究人员开发了一种新的方法来研究蛋白质复合体的组合. 他们发现酵母中的核孔综合体 (NPC) 形成了层次结构,其子综合体首先组合起来,然后在一个小时内成熟.

科学领域:

细胞生物学
生物化学
结构生物学

背景情况:

蛋白质复合体对于细胞功能至关重要.
描述像核孔复合体 (NPC) 这样的大型蛋白质复合体的组装动态仍然具有挑战性.
了解蛋白质复合组合是解读细胞过程的关键.

研究的目的:

为分析蛋白质复合体的原生组装动力学制定高通量策略.
描述酵母NPC中的核波林 (NUP) 的联合组装动态.
揭示NPC生物发生和蛋白质复合体组合的原理.

主要方法:

开发了一个高通量策略来分析本地装配动力学.
应用该方法研究酵母中320个核对的联合组合.
研究了核孔复合体的时间尺度和层次组合.

主要成果:

鉴定了核波林之间的不同联合组合率,从快速交换到漫长的成熟.
揭示了一个层次的NPC生物生成模型:分组组建 (分钟) 接着是中心到外围组装 (∼1小时).
发现Mlp1 (一种核素) 晚期结合,首选与成熟的NPC结合.

结论:

开发的高吞吐量策略可以分析蛋白质复合体组装动态.
NPC组装遵循一个分层的,时间依赖的过程.
这些发现为大细胞机械的动态组合提供了洞察力,并可应用于其他多蛋白组合.

关键词:

DIA质谱测试国际农业协会-MFA 美国组装顺序数据独立获取大分子复合生物发生成熟动力学代谢标签核孔复合体脉冲-SILAC AP-MS 的时间

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