蛋氨酸对奶牛乳腺上皮细胞亚细胞蛋白质组的影响研究

Influence of Methionine on Subcellular Proteins of Dairy Cow Mammary Gland Epithelial Cell

为寻找奶牛乳腺上皮细胞亚细胞结构中与泌乳相关的重要蛋白质、从蛋白质水平揭示乳蛋白合成的调控机制,应用双向凝胶电泳技术分离体外培养的蛋氨酸处理组与正常组奶牛乳腺上皮细胞蛋白质,利用Image Maser 2D软件对图谱进行对比分析,基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)和数据库检索鉴定,并采用实时荧光定量PCR技术在mRNA水平上验证2-DE结果。质谱鉴定出8个表达上调的差异蛋白质,大多数差异蛋白质的功能涉及细胞骨架构成、能量代谢等过程。这些差异点的发现为研究奶牛泌乳机理提供了有益的线索。

To understand the molecular mechanism at protein level and find the crucial proteins which correlate with lactating,two-dimensional electrophoresis was applied to separate subcellular proteins of healthy dairy cow mammary gland epithelial cells and methionine-supplemented dairy cow mammary gland epithelial cells cultured in vitro.The Image Master 2-DE software was used to characterize different proteins.By matrix assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF-MS） and database searching,proteins were identified.The 2-DE results were verified on the mRNA level by real time quantitative PCR.Eight differentially-expressed proteins from MS analysis were of up-regulated expression.The differentially displayed proteins between control and methionine-supplemented dairy cow mammary gland epithelial cells are useful for lactating mechanism research.