...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Science China Life Sciences >Toward a high resolution 2-DE profile of the normal human liver proteome using ultra-zoom gels
【24h】

Toward a high resolution 2-DE profile of the normal human liver proteome using ultra-zoom gels

机译：使用超变焦凝胶实现正常人肝蛋白质组的高分辨率2-DE谱

获取原文
   

获取外文期刊封面封底 >>

       
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The human liver is the largest organ in the body and has many important physiological functions. A global analysis of human liver proteins is essential for a better understanding of the molecular basis of the normal functions of the liver and of its diseases. As part of the Human Liver Proteome Project (HLPP), the goal of the present study was to visualize and detect as many proteins as possible in normal human livers using two-dimensional gel electrophoresis (2-DE). We have constructed a reference map of the proteins of human normal liver that can be used for the comprehensive analysis of the human liver proteome and other related research. To improve the resolution and enhance the detection of low abundance proteins, we developed and optimized narrow pH range ultra-zoom 2-DE gels. High resolution patterns of human liver in pH gradients 4.5–5.5, 5–6, 5.5–6.7, 6–9 and 6–11 are presented. To improve the poor resolution in the alkaline pH range of 2-DE gels, we optimized the isoelectric focusing protocol by including sample application using cup loading at the anode and incorporating 1.2% hydroxyethyl disulfide, 15% 2-propanol and 5% glycerol in the rehydration buffer. Using the optimized protocol, we obtained reproducibly better resolution in both analytical and preparative 2-DE gels. Compared with the 2386 and 1878 protein spots resolved in the wide range 3–10 and 4–7 pH gradients respectively, we obtained 5481 protein spots from the multiple (overlapping) narrow pH range ultra-zoom gels in the range of pH 4.5–9. The visualized reference map of normal human liver proteins presented in this paper will be valuable for comparative proteomic research of the liver proteome.
机译：人体肝脏是人体最大的器官，具有许多重要的生理功能。人类肝脏蛋白质的全局分析对于更好地了解肝脏及其疾病正常功能的分子基础至关重要。作为人类肝蛋白质组计划（HLPP）的一部分，本研究的目的是使用二维凝胶电泳（2-DE）可视化并检测正常人肝脏中尽可能多的蛋白质。我们已经构建了人类正常肝脏蛋白质的参考图谱，可用于人类肝脏蛋白质组学和其他相关研究的综合分析。为了提高分离度并增强对低丰度蛋白质的检测，我们开发并优化了窄pH范围超变焦2-DE凝胶。提出了人类肝脏在pH梯度4.5-5.5、5-6、5.5-6.7、6-9和6-11中的高分辨率模式。为了改善2-DE凝胶在碱性pH范围内的较差分离度，我们优化了等电点聚焦方案，包括在阳极上加样杯并在样品中加入1.2％羟乙基二硫化物，15％2-丙醇和5％甘油的样品应用。补液缓冲区。使用优化的方案，我们在分析型和制备型2-DE凝胶中均获得了可再现的更好分离度。与分别在3-10和4-7的宽pH梯度范围内分离的2386和1878的蛋白质点相比，我们从pH值为4.5-9的多个（重叠）窄pH范围超变焦凝胶中获得了5481的蛋白点。 。本文介绍的正常人肝蛋白的可视化参考图对于肝蛋白质组的比较蛋白质组学研究将是有价值的。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号