双向电泳分析蛋氨酸对奶牛乳腺上皮细胞核磷酸化蛋白质的影响

›› 2013, Vol. 40 ›› Issue (6): 53-57.

双向电泳分析蛋氨酸对奶牛乳腺上皮细胞核磷酸化蛋白质的影响

陆黎敏, 李庆章, 黄建国, 刘荣, 王佳丽, 金鑫, 高学军

东北农业大学, 乳品科学教育部重点实验室, 黑龙江哈尔滨 150030

收稿日期:2012-10-10 出版日期:2013-06-20 发布日期:2013-06-20
通讯作者: 高学军 E-mail:gaoxj5390@sina.com
作者简介:陆黎敏(1982-),女,黑龙江人,博士生,研究方向:蛋白质组学。
基金资助:
国家重点基础研究发展(973)计划(2011CB100804)。

Proteomic Analysis of the Nuclear Phosphorylated Proteins in Diary Cow Mammary Epithelial Cells Treated with Methionine

LU Li-min, LI Qing-zhang, HUANG Jian-guo, LIU Rong, WANG Jia-li, JIN Xin, GAO Xue-jun 

Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China

Received:2012-10-10 Online:2013-06-20 Published:2013-06-20

摘要/Abstract

摘要： 为了寻找奶牛乳腺上皮细胞核中与乳蛋白合成相关的重要蛋白质,从翻译水平揭示乳蛋白合成的机理,本试验应用双向凝胶电泳技术和基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)的方法,分析添加蛋氨酸后奶牛乳腺上皮细胞核磷酸化蛋白质的变化,并采用Western blotting验证。质谱分析鉴定出5个表达上调的细胞核磷酸化蛋白质,分别是葡萄球菌核酸域包含蛋白质1、甘氨酰-tRNA合成酶、Septin-6、Twinfilin-1和延长因子1-beta,这些蛋白质的功能涉及DNA转录、mRNA翻译、蛋白质合成、细胞分裂、细胞形态发生及细胞周期的调控。

关键词: 细胞核; 磷酸化蛋白质; 双向电泳; 奶牛; 乳腺上皮细胞

Abstract: To reveal the major protein related to milk protein synthesis in nucleus of mammary gland epithelial cells and further uncover the mechanism of milk protein synthesis in post-transcription level, two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization/time of flight mass spectrometry (MALDI-TOF-MS) were used to identify the changes of nuclear phosphorylated proteins in DCMECs treated with methionine, and their expression changes were verified by Western blotting analysis in the current study. 5 up-regulated expressed phosphoproteins included staphylococcal nuclease domain-containing protein 1, Septin-6, Glycyl tRNA synthetase, Twinfilin-1 and eukaryotic elongation factor 1-beta. They contributed to multiple functional activities such as DNA transcription, mRNA translation, protein synthesis, cell cycle, cell division and morphological processes.

Key words: nuclear; phosphorylated protein; 2-DE; diary cow; mammary epithelial cells

中图分类号:

S823

陆黎敏, 李庆章, 黄建国, 刘荣, 王佳丽, 金鑫, 高学军. 双向电泳分析蛋氨酸对奶牛乳腺上皮细胞核磷酸化蛋白质的影响[J]. , 2013, 40(6): 53-57.

LU Li-min, LI Qing-zhang, HUANG Jian-guo, LIU Rong, WANG Jia-li, JIN Xin, GAO Xue-jun . Proteomic Analysis of the Nuclear Phosphorylated Proteins in Diary Cow Mammary Epithelial Cells Treated with Methionine[J]. , 2013, 40(6): 53-57.

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