唐仙英,肖 瑶,海 力.蛋白体外结合实验联合质谱分析鉴定 Num1 的互作蛋白[J].中南民族大学学报自然科学版,2018,(4):27-29
蛋白体外结合实验联合质谱分析鉴定 Num1 的互作蛋白
Identification of Num1-interacting Proteins by Pull-down Experiments and Mass Spectrometry
  
DOI:10.12130/znmdzk.20180406
中文关键词: 芽殖酵母  动力蛋白  PA 结构域  纺锤体定位
英文关键词: budding yeast  dynein  PA domain  spindle orientation
基金项目:湖北省自然科学基金重点资助项目( 2011BMZ102)
作者单位
唐仙英,肖 瑶,海 力 中南民族大学 生命科学学院武陵山区特色资源植物种质保护与利用湖北省重点实验室武汉 430074 
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中文摘要:
      目的: 为了解析芽殖酵母 Num1 蛋白在纺锤体定位和线粒体中的调节机制.方法: 利用大肠杆菌原核表达 并纯化了在 Num1 功能中起核心作用的补丁组装(PA) 结构域的重组蛋白, 通过蛋白体外结合实验(Pull-down) 分 离了酵母细胞中与 PA 结构域相结合的蛋白复合物,并对其进行了质谱分析.结果: 鉴定了一系列新的 Num1 互作蛋 白,包括核糖体蛋白,参与蛋白折叠、分配及转运的内质网和高尔基复合体蛋白, 参与基因转录和翻译的核酸酶和 蛋白酶,及其他功能的蛋白.结论: Num1 的互作蛋白的鉴定为进一步研究 Num1 的作用机制奠定了基础.
英文摘要:
      Objective: To explore the regulatory mechanisms of Num1 during spindle orientation and in mitochondria in budding yeast. Method: Recombinant proteins containing the patch assembly (PA) domain, which played central roles in Num1's functions, were expressed and purified from E. coli cells. Protein complexes that bound to the PA domain were isolated from yeast cells through pull-down experiments and analyzed by mass spectrometry. Results: A series of novel Num1-interacting proteins were identified. These included ribosomal proteins, proteins involved in protein folding, sorting and transportation in the ER and Golgi complexes, some nucleases and protein enzymes involved in gene transcription and translation, and other proteins with various functions. Conclusion: Identification of Num1-interacting proteins lays a foundation for further study of Num1's action mechanism .
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