Ribotrap Analysis of Proteins Associated with FHL3 3’Untranslated Region in Glioma Cells

Wei Han; Qing Xia; Bin Yin Xiao-zhong Peng

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Ribotrap Analysis of Proteins Associated with FHL3 3’Untranslated Region in Glioma Cells

机译：胶质瘤细胞中与FHL3 3′非翻译区相关的蛋白质的核糖分析

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Objective To screen the proteins associated with four-and-a-half LIM domains 3 (FHL3) 3’ untranslated region (3’UTR) in glioma cells. Methods Western blot was adopted to detect the regulatory effect of poly(C)-binding protein 2 (PCBP2) on FHL3. Biotin pull-down and sliver staining were employed to screen and verify the candidate binding proteins of FHL3 3’UTR. Then liquid chromatography-tandem mass spectrometry (LC-MS/MS) and molecule annotation system were used to identify and analyze the candidate binding proteins. Immuno-precipitation was conducted to study the interaction between PCBP2 and polypyrimidine tract-binding protein 1 (PTBP1), a binding protein identified by LC-MS/MS. Results PCBP2 could bind to FHL3 mRNA 3’UTR-A and inhibited the expression of FHL3 in T98G glioms cells. 22 candidate binding proteins were identified. Among them, there were 11 RNA binding proteins, including PCBP2. PTBP1 associated with FHL3 mRNA 3’UTR and interacted with PCBP2 protein. Conclusion PCBP2 and PTBP1 can both associate with FHL3 mRNA 3’UTR through forming a protein complex.

机译：目的筛选与神经胶质瘤细胞中的四个半LIM结构域3（FHL3）3'非翻译区（3'UTR）相关的蛋白质。方法采用Western印迹法检测poly（C）结合蛋白2（PCBP2）对FHL3的调节作用。生物素下拉法和银染法用于筛选和验证FHL3 3’UTR的候选结合蛋白。然后使用液相色谱-串联质谱（LC-MS / MS）和分子注释系统鉴定和分析候选结合蛋白。进行了免疫沉淀研究PCBP2和多嘧啶束结合蛋白1（PTBP1）之间的相互作用，该蛋白通过LC-MS / MS鉴定。结果PCBP2可以与FHL3 mRNA 3’UTR-A结合，并抑制T98G胶质瘤细胞中FHL3的表达。鉴定了22种候选结合蛋白。其中，有11种RNA结合蛋白，包括PCBP2。 PTBP1与FHL3 mRNA 3’UTR相关，并与PCBP2蛋白相互作用。结论结论PCBP2和PTBP1均可通过形成蛋白质复合物而与FHL3 mRNA 3’UTR结合。

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《中国医学科学杂志（英文版）》 |2014年第2期|78-84|共7页
作者
Wei Han; Qing Xia; Bin Yin Xiao-zhong Peng;
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State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences&Peking Union Medical College, Beijing 100005, China;

State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, Beijing 100850, China;

State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences&Peking Union Medical College, Beijing 100005, China;

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收录信息中国科学引文数据库(CSCD);中国科技论文与引文数据库(CSTPCD);
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正文语种 eng
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Ribotrap Analysis of Proteins Associated with FHL3 3’Untranslated Region in Glioma Cells

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