通过一步萃取将组蛋白分为多个亚组分的蛋白质组预分离方法的建立

高等学校化学学报 ›› 2010, Vol. 31 ›› Issue (7): 1314.

通过一步萃取将组蛋白分为多个亚组分的蛋白质组预分离方法的建立

白海鑫¹, 刘小花¹, 杨帆², 杨秀荣², 汪尔康²

1. 河南农业大学理学院应用化学系, 郑州 450002;
2. 中国科学院长春应用化学研究所电分析化学国家重点实验室, 长春 130022

收稿日期:2009-12-28 出版日期:2010-07-10 发布日期:2010-07-10
通讯作者: 汪尔康, 男, 研究员, 博士生导师, 中国科学院院士, 主要从事电分析化学研究. E-mail: ekwang@ciac.jl.cn; 白海鑫, 男, 博士, 讲师, 主要从事生物分析及材料化学研究. E-mail: haixin-bai@yahoo.com.cn
基金资助:
国家自然科学基金(批准号: 20735003)和电分析化学国家重点实验室基金(批准号: SKLEAC2010011)资助.

Prefractionation Method to Separate Proteomic Proteins into Multigroups by One-step Extraction

BAI Hai-Xin^1*, LIU Xiao-Hua¹, YANG Fan², YANG Xiu-Rong², WANG Er-Kang^2*

1. Department of Applied Chemistry, College of Sciences, Henan Agricultural University, Zhengzhou 450002, China;
2. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China

Received:2009-12-28 Online:2010-07-10 Published:2010-07-10
Contact: WANG Er-Kang. E-mail: ekwang@ciac.jl.cn; BAI Hai-Xin. E-mail: haixin-bai@yahoo.com.cn
Supported by:
国家自然科学基金(批准号: 20735003)和电分析化学国家重点实验室基金(批准号: SKLEAC2010011)资助.

摘要/Abstract

摘要：

将基于正丙醇、氯化钠和水的双水相体系用于蛋白质组的预分离研究, 并考察了体系酸度及在不同浓度的盐、正丙醇及蛋白质存在时该双水相体系对蛋白质的分离效果. 将经该双水相体系预分离过的蛋白质组样品在未与成相试剂分离的条件下直接用于凝胶电泳分析. 结果表明, 该双水相体系可通过一步萃取将蛋白质组样品分为3个亚组群. 该蛋白质组预分离方法简单、快速、成本低, 并具有生物相容性、可连续操作性、无需昂贵复杂仪器以及在进行电泳分析前无须(或易于)将目标蛋白与成相试剂分离等优点. 该蛋白质组预分离方法的建立在蛋白质组学和方法学方面均有着极为重要的意义.

关键词: 蛋白质组预分离方法; 双水相体系; 萃取; 分离

Abstract:

For the first time, an aqueous two-phase system(ATPS) composed of n-propanol, NaCl and water was applied in prefractionating proteomic proteins. In this work, ethanol, n-propanol and several commonly used inorganic salts were attempted to develop the ATPS. The ATPS composed of n-propanol, NaCl and water was developed finally. Subsequently, the effects of pH, salt concentration, n-propanol volume and protein concentration on the ATPS′ separation effectiveness for proteins were studied. Protein samples were prefractionated by the ATPS and then analyzed by gel electrophoresis. The results proved that the prefractionation method could separate proteomic proteins into three groups by one-step extraction, and there was no need for separating target proteins from the phase-forming reagents prior to gel electrophoresis analysis. Simplicity, rapidity and low cost are the highlight attractiveness of the prefractionation method. It also has advantages of biocompability, potential for continuous operation, no requirement for sophisticated instrumentations and being easy or no need for separating the target proteins from phase-forming reagents prior to electrophoresis analysis. The prefractionation method has great significance in proteomics and methodology.

Key words: Proteomic prefractionation method; Aqueous two-phase system; Extraction; Separation

TrendMD:

白海鑫, 刘小花, 杨帆, 杨秀荣, 汪尔康. 通过一步萃取将组蛋白分为多个亚组分的蛋白质组预分离方法的建立. 高等学校化学学报, 2010, 31(7): 1314.

BAI Hai-Xin*, LIU Xiao-Hua, YANG Fan, YANG Xiu-Rong, WANG Er-Kang*. Prefractionation Method to Separate Proteomic Proteins into Multigroups by One-step Extraction. Chem. J. Chinese Universities, 2010, 31(7): 1314.

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