Novel interacting proteins identified by tandem affinity purification coupled to nano LC-MS/MS interact with ribosomal S6 protein kinase 4 (RSK4) and its variant protein (RSK4m)
Novel interacting proteins identified by tandem affinity purification coupled to nano LC-MS/MS interact with ribosomal S6 protein kinase 4 (RSK4) and its variant protein (RSK4m)
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DOI：
10.1016/j.ijbiomac.2016.12.030
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发表时间：
2017-03-01
影响因子：
8.2
通讯作者：
Yang, Huawei
中科院分区：
文献类型：
--
作者：
Liu, Riqiang;Liu, Tianhua;Yang, Huawei
核糖体S6蛋白激酶4(RSK4)是一种重要的新型肿瘤抑制因子，具有抑制乳腺癌细胞生长和诱导衰老的作用。然而，RSK4m是由选择性剪接产生的RSK4的变体，可能在某些方面发挥不同的作用。关于RSK4或RSK4m活性的机制一直缺乏了解。通过对RSK4和RSK4m相互作用组的分析，可以深入了解它们的特定功能和整合机制。通过串联亲和纯化，获得了与RSK4或RSK4m相互作用的蛋白质复合体。对得到的蛋白质复合体进行了质谱鉴定，并进行了生物信息学分析。在这项研究中，我们利用两个链球菌II和一个基于单标志标签的串联亲和纯化技术(SF-TAP)结合纳米LC-MS/MS在MDA-MB-231细胞中分离和鉴定了82个RSK4相关蛋白和137个RSK4m相关蛋白。基因本体论和独创性路径分析为RSK4和RSK4m的蛋白质相互作用组提供了功能注释和蛋白质-蛋白质相互作用网络。通过生物信息学分析对这些蛋白质的功能注释强调了RSK4和RSK4m与其相互作用的伙伴协调在调节多种生物过程中的重要作用，特别是细胞衰老。此外，通过比较RSK4和RSK4m的相互作用组，我们发现RSK4m比RSK4参与了更多的分子功能。(C)2016年提交人。由Elsevier B.V.出版。这是CC BY-NC-ND许可证(http://creativecommons.org/licenses/by-nc-nd/4.0/).下的一篇开放获取文章
Ribosomal S6 protein kinase 4 (RSK4) is an important novel tumor suppressor that inhibits breast cancer cell growth and induces senescence. However, RSK4m, which is a variant of RSK4 resulting from alternative splicing, may play distinct roles in some aspects. Knowledge about the mechanisms of RSK4 or RSK4m activity has been lacking. Analysis of the RSK4 and RSK4m interactome could provide insight into their specific functions and integrative mechanisms. Using tandem affinity purification, we obtained protein complexes that interacted with RSK4 or RSK4m. Mass spectrum analysis was performed to identify the obtained protein complexes, and bioinformatics analysis was performed. In this study, we isolated and identified 82 RSK4-associated proteins and 137 RSK4m-associated proteins using two STREP II and a single Flag tag-based tandem affinity purification (SF-TAP) coupled with nano LC-MS/MS in MDA-MB-231 cells. Gene Ontology and Ingenuity Pathway Analysis analyses provided functional annotations and protein-protein interaction networks of the protein interactome of RSK4 and RSK4m. Functional annotations of these proteins by bioinformatics analyses highlight the essential role of RSK4 and RSK4m in coordination with their interacting partners to mediate multiple biological processes, especially cell senescence. Moreover, after comparing the interactome of RSK4 and RSK4m, we found that RSK4m is involved in more molecular functions than RSK4. (C) 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).