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液相等电聚焦结合双向凝胶电泳分离碱性蛋白
引用本文:钟华,张扬,樊惠芝,杨芃原. 液相等电聚焦结合双向凝胶电泳分离碱性蛋白[J]. 高等学校化学学报, 2006, 27(10): 1830-1834
作者姓名:钟华  张扬  樊惠芝  杨芃原
作者单位:复旦大学生物医学研究院,蛋白质组学研究中心,上海,200433
基金项目:国家自然科学基金(批准号:90408023),国家科技攻关计划(批准号:2004BA711A18),上海市重大基础研究项目(批准号:04DZ14005)资助.
摘    要:在蛋白组学研究中, 经典的双向凝胶电泳法(2-DE)对碱性蛋白及低丰度蛋白的分离存在技术障碍, 但预分离技术的应用可弥补其缺陷. 液相等电聚焦可有效地分离富集复杂蛋白样品. 碱性胶条用于2-DE可极大地提高蛋白上样量和凝胶分辨率. 将上述两种技术相结合用于碱性蛋白质和低丰度蛋白质的分离鉴定, 可使碱端区域双向凝胶图谱质量显著提高, 蛋白点更清晰且点数增多, 质谱鉴定确信度提高, 碱性蛋白和低丰度蛋白质谱鉴定成功率提高, 对于蛋白组学研究具有一定的意义.

关 键 词:液相等电聚焦  双向凝胶电泳  碱性蛋白
文章编号:0251-0790(2006)10-1830-05
收稿时间:2006-04-10
修稿时间:2006-04-10

Combination of Liquid Isoelectric Focusing and 2-DE Applied to the Analysis of Alkaline Proteins
ZHONG Hua,ZHANG Yang,FAN Hui-Zhi,YANG Peng-Yuan. Combination of Liquid Isoelectric Focusing and 2-DE Applied to the Analysis of Alkaline Proteins[J]. Chemical Research In Chinese Universities, 2006, 27(10): 1830-1834
Authors:ZHONG Hua  ZHANG Yang  FAN Hui-Zhi  YANG Peng-Yuan
Affiliation:Proteome Center, Academy of Biomedicine, Fudan University, Shanghai 200433, China
Abstract:Alkaline proteins which represent a large part of proteome,remain a great challenge for conventional 2-DE.Fractionation methods can offset the weakness of 2-DE for the analysis of these proteins.In this work,liquid isoelectric focusing(LIEF)and two-dimensional electrophoresis(2-DE)were conbined for the analysis of alkaline proteins from rat liver.Under this strategy,the resolution of alkaline proteins on 2-D gel and reliability of proteins identification by MS were greatly improved,and more low-abundance and highly alkaline proteins were successfully identified compared to 2-DE without LIEF.
Keywords:Liquid isoelectric focusing  Two-dimensional electrophoresis  Alkaline protein
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