Department of Biotechnology, Delft University of Technology, Julianalaan 67, NL-2628 BC Delft.
Chem Biodivers. 2012 Sep;9(9):1967-80. doi: 10.1002/cbdv.201100412.
Metalloproteomics is a rapidly developing field of science that involves the comprehensive analysis of all metal-containing or metal-binding proteins in a biological sample. The purpose of this review is to offer an overview of the research involving Metal Isotope native RadioAutography in Gel Electrophoresis (MIRAGE), a powerful new method to visualize and study the proteome of a particular metal ion. MIRAGE involves four steps: i) labelling of target proteins with a radioisotope; ii) separation of intact holo-proteins using native isoelectric focusing (1D) combined with Blue Native PAGE (2D); iii) spot visualization and quantification using autoradiography; and iv) protein identification by tandem mass spectrometry. MIRAGE Investigations of the soluble Cu, Zn, and Fe metalloproteomes of Escherichia coli, and of the soluble Mo and W proteomes of the hyperthermophilic archaeon Pyrococcus furiosus are reviewed.
金属蛋白质组学是一个快速发展的科学领域，涉及对生物样本中所有含金属或与金属结合的蛋白质进行全面分析。本文综述了金属同位素天然放射自显影凝胶电泳（MIRAGE）的研究进展，这是一种可视化和研究特定金属离子蛋白质组的强大新方法。MIRAGE 实验包括四个步骤：i）用放射性同位素标记靶蛋白；ii）使用天然等电聚焦（1D）结合蓝色天然聚丙烯酰胺凝胶电泳（2D）分离完整的全蛋白；iii）使用放射自显影进行斑点可视化和定量；iv）通过串联质谱进行蛋白质鉴定。本文还综述了大肠杆菌可溶性 Cu、Zn 和 Fe 金属蛋白质组学以及嗜热古菌 Pyrococcus furiosus 可溶性 Mo 和 W 蛋白质组学的 MIRAGE 研究。
