| 丁 燕,郑子萌,赵婷婷,石清华,徐宇君.哺乳动物PUF家族蛋白PUMILIO1的伴侣蛋白研究[J].,2017,17(19):3606-3609 |
| 哺乳动物PUF家族蛋白PUMILIO1的伴侣蛋白研究 |
| Identification of a PUMILIO1 Associated Protein in Mammalian Cells |
| 投稿时间：2017-02-12 修订日期：2017-02-28 |
| DOI：10.13241/j.cnki.pmb.2017.19.002 |
| 中文关键词: PUF PUM1 NDFIP2 NIH3T3细胞 DU145 细胞 |
| 英文关键词: PUF PUM1 NDFIP2 NIH3T3 cells DU145 cells |
| 基金项目:国家自然科学基金面上项目(81270737)；江苏省自然科学基金面上项目(BK2012838) |
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| 中文摘要: |
| 摘要 目的：探索NIH3T3和DU145细胞中PUM1蛋白的伴侣蛋白。方法：收集NIH3T3和DU145细胞进行免疫沉淀实验，实验组和对照组分别使用抗PUM1抗体和抗Goat-IgG抗体。通过银染实验观察实验组与对照组是否有差异性条带，通过质谱鉴定和Mascot软件对差异性条带所包含的蛋白进行分析，鉴定PUM1蛋白在细胞中发挥作用时的可能互动蛋白。结果：在NIH3T3和DU145细胞银染实验中，实验组凝胶泳道均在约37 KD处出现差异性条带。对NIH3T3细胞中差异性条带进行质谱分析，通过Mascot软件肽质量指纹谱搜索，将有真实差异的蛋白评分最低阈值设为65，P<0.05，结果显示肽段评分最高的蛋白为NDFIP2蛋白。与NCBInr数据库比对，检测到针对NDFIP2的肽段的覆盖率为40 %。结论：NDFIP2与哺乳动物PUM1蛋白在细胞内关联，可能是PUF家族蛋白的伴侣蛋白。 |
| 英文摘要: |
| ABSTRACT Objective: To explore PUMILIO1 (PUM1) associated proteins in mammalian cells (NIH3T3 and DU145 cells). Methods: NIH3T3 and DU145 cells were respectively collected. Proteins were obtained by IP (Immunoprecipitation) experiment. Experimental group and control group were set up using anti PUM1 antibody and anti Goat-IgG antibody, respectively. Silver staining revealed differ- ential bands in PUM1 pull-down lane in comparison to IgG lane. Proteins from the differential bands were analyzed by mass spectrum and Mascot software analysis. Results: A macroscopic differential band around 37 KD in experimental group was observed both in NIH3T3 and DU145 cells. NDFIP2 protein acquired the highest Mascot score using mass spectrum and peptide mass fingerprint in which the minimum threshold value of the score representing real differences of protein was set to 65(P<0.05). The peptides coverage specific to NDFIP2 was forty percent in reference to NCBInr database. Conclusion: NDFIP2 was associated with PUM1 protein in mammalian cells and might be a partner protein of PUF family proteins. |
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