Structural dynamics of the human COP9 signalosome revealed by cross-linking mass spectrometry and integrative modeling
Structural dynamics of the human COP9 signalosome revealed by cross-linking mass spectrometry and integrative modeling
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DOI：
10.1073/pnas.1915542117
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发表时间：
2020-02-25
影响因子：
11.1
通讯作者：
Huang, Lan
中科院分区：
文献类型：
--
作者：
Gutierrez, Craig;Chemmama, Ilan E.;Huang, Lan
COP 9信号体（CSN）是一种进化上保守的八亚基（CSN 1 -8）蛋白复合物，其通过去甲基化Cullin-RING E3连接酶（CRL 5）控制蛋白泛素化。CSN的激活和功能取决于其结构动力学，这一直是具有挑战性的破译传统工具。在这里，我们开发了一种多化学交联质谱法，通过三种质谱可裂解的交联剂来生成高度可靠的交联数据。我们应用这种方法与综合结构建模，以确定CSN的相互作用和结构动力学与最近发现的第九亚基，CSN 9，在解决方案。我们的研究结果确定了CSN 9结合位点的定位，并揭示了CSN 9依赖的CSN结构变化。结合生化分析，我们提出了一个结构模型，其中CSN 9结合触发CSN采取的配置，有利于CSN-CRL相互作用，从而增强CSN脱羧酶活性。我们的综合结构分析工作流程可以概括为定义其他方法无法访问的动态蛋白质复合物的溶液内架构。
The COP9 signalosome (CSN) is an evolutionarily conserved eight-subunit (CSN1-8) protein complex that controls protein ubiquitination by deneddylating Cullin-RING E3 ligases (CRL5). The activation and function of CSN hinges on its structural dynamics, which has been challenging to decipher by conventional tools. Here, we have developed a multichemistry cross-linking mass spectrometry approach enabled by three mass spectometry-cleavable cross-linkers to generate highly reliable cross-link data. We applied this approach with integrative structure modeling to determine the interaction and structural dynamics of CSN with the recently discovered ninth subunit, CSN9, in solution. Our results determined the localization of CSN9 binding sites and revealed CSN9-dependent structural changes of CSN. Together with biochemical analysis, we propose a structural model in which CSN9 binding triggers CSN to adopt a configuration that facilitates CSN-CRL interactions, thereby augmenting CSN deneddylase activity. Our integrative structure analysis workflow can be generalized to define in-solution architectures of dynamic protein complexes that remain inaccessible to other approaches.