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

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.

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

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.