微柱高效液相色谱-质谱/质谱快速鉴定混合蛋白质新方法

A New Technique to Identify Individual Protein in Complex by Microcolumn-HPLC-ESIMS/MS

发展了一种混合蛋白质快速鉴定的新方法 .将几种蛋白质的混合物于热变性后直接在溶液中酶解 ,利用微柱高效液相色谱 -离子阱串级质谱进行肽谱 /氨基酸序列分析 ,并结合 Mascot数据库搜索处理功能 。

In this work the identification method for protein mixture is reported, in which several proteins were varified by HPLC ESIMS/MS after the whole complex proteins were directly digested in solution, instead of the routine 2 dimensional gel electrophoresis. The obtained data was then searched against NCBInr database by using MS/MS Ions Search(http://www.matrixscience.com), and a positive result was therefore obtained. With the simulated sequence spectra in database searching, a re identification method was developed for low score matched protein by sequence tag technique. As a typical example, Concanavalin A, a less soluble protein with a low matching score, was also successfully identified, which indicated the great potential of complex digestion in proteomic study.