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首页> 外文会议>4th international symposium on dairy cow nutrition and milk quality proceedings >A cow mammary epithelial cell-free system based on crude lysosomes and cytosol proteins:leucine activating mTOR at Ser2448
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A cow mammary epithelial cell-free system based on crude lysosomes and cytosol proteins:leucine activating mTOR at Ser2448

机译：基于粗制溶酶体和胞浆蛋白的牛乳腺上皮无细胞系统：Ser2448上的亮氨酸激活mTOR

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摘要

Essential amino acids,especially leucine,initiated the signaling of the mammalian target of rapamycin complex-1 (mTORC1) and protein synthesis in cow mammary epithelial cells(CMEC).CMEC-H is an immortalized cow mammary epithelial cell line.We established a cell-free system,which was based on crude lysosomes and cytosol proteins from CMEC-H.However,the model can partially duplicate aspects of amino acids signaling mTOR program in vitro.The addition of l×leucine solely and 1 ×essential amino acids led mTOR to be activated,phosphorylated and then moved towards the crude lysosome fractions in the cell-free system.However,combined with 1 ×leucine or 1 ×essential amino acids,20 nm rapamycin was not able to completely prevent mTOR from being phosphorylated.By trypan blue dye exclusion assay,we found passing through gauge-26 needle was a good cell lysis method of CMEC-H.This study may provide some directions for further construction of cell-free system duplicating amino acid signaling mTOR.Futhurmore,the method may apply to cell-free models of other cell lines.
机译：必需氨基酸，特别是亮氨酸，引发了哺乳动物乳腺上皮细胞（CMEC）中雷帕霉素复合物-1（mTORC1）的信号转导和蛋白质合成。CMEC-H是永生化的乳腺上皮细胞系。我们建立了一个细胞该系统是基于CMEC-H的粗溶酶体和胞质蛋白的无系统系统。但是，该模型可以部分复制体外信号转导mTOR程序的氨基酸。单独添加1×亮氨酸和1×必需氨基酸可导致mTOR。被激活，磷酸化然后移向无细胞系统中的粗溶酶体部分。但是，与1×亮氨酸或1×必需氨基酸结合使用时，20 nm雷帕霉素不能完全阻止mTOR磷酸化。蓝色染料排斥实验，发现通过26号针头是CMEC-H的一种良好的细胞裂解方法。这项研究可能为进一步构建复制氨基酸信号mTOR的无细胞系统提供一些指导。此外，该方法可能适用于其他细胞系的无细胞模型。

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  • 来源
  • 会议地点 Beijing(CN)
  • 作者

    W.T.Dai; H.Hu; J.Q.Wang; N.Zheng;

  • 作者单位

    Ministry of Agriculture Laboratory of Quality Safety Risk Assessment for Dairy Products(Beijing),Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, P.R.China;

    Ministry of Agriculture Laboratory of Quality Safety Risk Assessment for Dairy Products(Beijing),Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, P.R.China;

    Ministry of Agriculture Laboratory of Quality Safety Risk Assessment for Dairy Products(Beijing),Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, P.R.China;

    Ministry of Agriculture Laboratory of Quality Safety Risk Assessment for Dairy Products(Beijing),Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, P.R.China;

  • 会议组织
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 乳产品综合利用;
  • 关键词

    Cell-free system; Crude Lysosomes; Leucine; mTOR; Cow mammary epithelial cell;

    机译：无细胞系统；粗制溶酶体；亮氨酸； mTOR；牛乳腺上皮细胞;

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