Structural Probing of a Protein Phosphatase 2A Network by Chemical Cross-Linking and Mass Spectrometry
Structural Probing of a Protein Phosphatase 2A Network by Chemical Cross-Linking and Mass Spectrometry
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DOI：
10.1126/science.1221483
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发表时间：
2012-09-14
期刊：
影响因子：
56.9
通讯作者：
Aebersold, Ruedi
中科院分区：
文献类型：
--
作者：
Herzog, Franz;Kahraman, Abdullah;Aebersold, Ruedi
通过化学交联和质谱法（XL-MS）鉴定近似氨基酸有助于同质蛋白质复合物的结构分析。我们获得了距离限制的模块化相互作用网络的蛋白质复合物亲和纯化从人类细胞通过应用一个适应的XL-MS协议。对人蛋白磷酸酶2A（PP 2A）复合物的系统分析确定了176个蛋白质间和570个蛋白质内交联，这些交联将特定的三聚体PP 2A复合物连接到控制其细胞功能的众多衔接蛋白。空间限制引导免疫球蛋白结合蛋白1（IGBP 1）和PP 2A之间的结合界面的分子建模，并揭示了TCP 1环复合物（TRiC）伴侣蛋白与PP 2A调节亚基2ABG相互作用的拓扑结构。本研究建立了XL-MS作为内源性蛋白质复合物分析的混合结构生物学方法的一个组成部分。
The identification of proximate amino acids by chemical cross-linking and mass spectrometry (XL-MS) facilitates the structural analysis of homogeneous protein complexes. We gained distance restraints on a modular interaction network of protein complexes affinity-purified from human cells by applying an adapted XL-MS protocol. Systematic analysis of human protein phosphatase 2A (PP2A) complexes identified 176 interprotein and 570 intraprotein cross-links that link specific trimeric PP2A complexes to a multitude of adaptor proteins that control their cellular functions. Spatial restraints guided molecular modeling of the binding interface between immunoglobulin binding protein 1 (IGBP1) and PP2A and revealed the topology of TCP1 ring complex (TRiC) chaperonin interacting with the PP2A regulatory subunit 2ABG. This study establishes XL-MS as an integral part of hybrid structural biology approaches for the analysis of endogenous protein complexes.