鸟枪法蛋白质组学：一种应用固定化pH梯度等电聚焦和液相色谱-串联质谱的定性方法。

Shotgun proteomics: a qualitative approach applying isoelectric focusing on immobilized pH gradient and LC-MS/MS.

作者信息

Geiser Laurent, Vaezzadeh Ali R, Deshusses Jacques M P, Hochstrasser Denis F

机构信息

Swiss Centre for Applied Human Toxicology, Geneva University, Geneva, Switzerland.

出版信息

Methods Mol Biol. 2011;681:449-58. doi: 10.1007/978-1-60761-913-0_26.

DOI:10.1007/978-1-60761-913-0_26

PMID:20978982

Abstract

Shotgun proteomics is a rapid and near universal strategy to identify proteins in complex protein mixtures. After protein digestion, the resulting peptide mixture is submitted to two orthogonal techniques: peptides are first separated according to their isoelectric point (pI) by isoelectric focusing (IEF) on immobilized pH gradient (IPG); after peptide extraction, they are then separated in the second dimension according to their hydrophobic properties by reverse phase liquid chromatography (RPLC). Finally, they are detected by tandem mass spectrometry (MS/MS) and proteins are matched by means of bioinformatics software.

摘要

鸟枪法蛋白质组学是一种快速且几乎通用的策略，用于鉴定复杂蛋白质混合物中的蛋白质。蛋白质消化后，所得肽混合物会采用两种正交技术：首先通过固定化pH梯度（IPG）上的等电聚焦（IEF）根据肽的等电点（pI）进行分离；肽提取后，再通过反相液相色谱（RPLC）根据其疏水性质在第二维中进行分离。最后，通过串联质谱（MS/MS）进行检测，并借助生物信息学软件对蛋白质进行匹配。