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首页> 外文期刊>Molecules >Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis
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Proteomic and Functional Analyses Reveal MAPK1 Regulates Milk Protein Synthesis

机译：蛋白质组学和功能分析揭示MAPK1调节乳蛋白合成

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L-Lysine (L-Lys) is an essential amino acid that plays fundamental roles in protein synthesis. Many nuclear phosphorylated proteins such as Stat5 and mTOR regulate milk protein synthesis. However, the details of milk protein synthesis control at the transcript and translational levels are not well known. In this current study, a two-dimensional gel electrophoresis (2-DE)/MS-based proteomic technology was used to identify phosphoproteins responsible for milk protein synthesis in dairy cow mammary epithelial cells (DCMECs). The effect of L-Lys on DCMECs was analyzed by CASY technology and reversed phase high performance liquid chromatography (RP-HPLC). The results showed that cell proliferation ability and β-casein expression were enhanced in DCMECs treated with L-Lys. By phosphoproteomics analysis, six proteins, including MAPK1, were identified up-expressed in DCMECs treated with 1.2 mM L-Lys for 24 h, and were verified by quantitative real-time PCR (qRT-PCR) and western blot. Overexpression and siRNA inhibition of MAPK1 experiments showed that MAPK1 upregulated milk protein synthesis through Stat5 and mTOR pathway. These findings that MAPK1 involves in regulation of milk synthesis shed new insights for understanding the mechanisms of milk protein synthesis.
机译：L-赖氨酸（L-Lys）是一种必需氨基酸，在蛋白质合成中起基本作用。许多核磷酸化蛋白（例如Stat5和mTOR）调节牛奶蛋白的合成。然而，在转录和翻译水平上乳蛋白合成控制的细节尚不清楚。在当前的这项研究中，基于二维凝胶电泳（2-DE）/ MS的蛋白质组学技术被用来鉴定负责奶牛乳腺上皮细胞（DCMEC）中牛奶蛋白合成的磷蛋白。通过CASY技术和反相高效液相色谱（RP-HPLC）分析了L-Lys对DCMEC的影响。结果显示，用L-Lys处理的DCMEC中，细胞增殖能力和β-酪蛋白表达增强。通过磷酸化蛋白质组学分析，确定了在用1.2 mM L-Lys处理24 h的DCMEC中高表达的6种蛋白质，包括MAPK1，并通过定量实时PCR（qRT-PCR）和蛋白质印迹进行了验证。 MAPK1的过表达和siRNA抑制实验表明MAPK1通过Stat5和mTOR途径上调了乳蛋白的合成。 MAPK1参与调节牛奶合成的这些发现为了解牛奶蛋白质合成的机理提供了新的见识。

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