Unraveling the surface glycoprotein interaction network by integrating chemical crosslinking with MS-based proteomics.
Unraveling the surface glycoprotein interaction network by integrating chemical crosslinking with MS-based proteomics.
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通过将化学交联与基于MS的蛋白质组学集成，从而揭示表面糖蛋白相互作用网络。
DOI：
10.1039/d0sc06327d
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发表时间：
2021-01-04
期刊：
影响因子：
8.4
通讯作者：
Wu R
中科院分区：
文献类型：
--
作者：
Sun F;Suttapitugsakul S;Wu R
细胞质膜为细胞内外的信息传递提供了高度交互的平台。表面糖蛋白相互作用网络在许多细胞外事件中极其重要，异常的蛋白质相互作用与包括癌症在内的多种疾病密切相关。对细胞表面蛋白相互作用的全面分析将加深我们对表面蛋白之间的协作以调节细胞活动的理解。在这项工作中，我们开发了一种结合化学交联、酶促反应和基于质谱的蛋白质组学的方法，系统地表征与表面糖蛋白相互作用的蛋白质，然后构建表面组相互作用网络。与蛋白质共价结合的聚糖被用作“诱饵”，与表面糖蛋白相互作用的蛋白质通过化学交联连接。表面糖蛋白上的聚糖被半乳糖氧化酶（GAO）氧化，随后通过酰肼化学富集表面糖蛋白及其相互作用物（“猎物”）。结合定量蛋白质组学，鉴定了超过 300 种与表面糖蛋白相互作用的蛋白质。在这些蛋白质上发现了许多与细胞外事件相关的重要结构域。基于蛋白质-蛋白质相互作用数据库，我们构建了已识别蛋白质之间的相互作用网络，其中枢纽蛋白在相互作用组中发挥更重要的作用。通过分析交联肽，鉴定出细胞表面糖蛋白的特定相互作用物。新开发的方法可广泛应用于研究细胞表面糖蛋白相互作用，包括细胞表面组与外部刺激相互作用的动力学。通过整合化学交联、酶氧化和基于 MS 的蛋白质组学，系统地表征了与细胞表面糖蛋白相互作用的蛋白质。然后构建表面糖蛋白相互作用网络。
The cell plasma membrane provides a highly interactive platform for the information transfer between the inside and outside of cells. The surface glycoprotein interaction network is extremely important in many extracellular events, and aberrant protein interactions are closely correlated with various diseases including cancer. Comprehensive analysis of cell surface protein interactions will deepen our understanding of the collaborations among surface proteins to regulate cellular activity. In this work, we developed a method integrating chemical crosslinking, an enzymatic reaction, and MS-based proteomics to systematically characterize proteins interacting with surface glycoproteins, and then constructed the surfaceome interaction network. Glycans covalently bound to proteins were employed as “baits”, and proteins that interact with surface glycoproteins were connected using chemical crosslinking. Glycans on surface glycoproteins were oxidized with galactose oxidase (GAO) and sequentially surface glycoproteins together with their interactors (“prey”) were enriched through hydrazide chemistry. In combination with quantitative proteomics, over 300 proteins interacting with surface glycoproteins were identified. Many important domains related to extracellular events were found on these proteins. Based on the protein–protein interaction database, we constructed the interaction network among the identified proteins, in which the hub proteins play more important roles in the interactome. Through analysis of crosslinked peptides, specific interactors were identified for glycoproteins on the cell surface. The newly developed method can be extensively applied to study glycoprotein interactions on the cell surface, including the dynamics of the surfaceome interactions in cells with external stimuli. Proteins interacting with glycoproteins on the cell surface were systematically characterized by integrating chemical crosslinking, enzymatic oxidation, and MS-based proteomics. The surface glycoprotein interaction network was then constructed.
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影响因子：
15.3
作者：
Bonifaz, L;Bonnyay, D;Mahnke, K;Rivera, M;Nussenzweig, MC;Steinman, RM
通讯作者：
Steinman, RM
影响因子：
64.5
作者：
Dennis JW;Nabi IR;Demetriou M
通讯作者：
Demetriou M
影响因子：
5.6
作者：
Corgiat BA;Nordman JC;Kabbani N
通讯作者：
Kabbani N
DOI：
10.1084/jem.182.2.419
发表时间：
1995-08-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Katoh S;Zheng Z;Oritani K;Shimozato T;Kincade PW
通讯作者：
Kincade PW
影响因子：
14.8
作者：
Han, S;Collins, BE;Paulson, JC
通讯作者：
Paulson, JC