Multidimension polyacrylamide gel electrophoresis oriented toward the identification of metalloproteins

2015 Fiscal Year Final Research Report

• Project/Area Number: 25410139
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Analytical chemistry
• Research Institution: Saitama University
• Principal Investigator: SAITO Shingo 埼玉大学, 理工学研究科, 准教授 (60343018)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: メタロミクス / 金属タンパク質 / 電気泳動 / スペシエーション

Outline of Final Research Achievements

Although investigation of metalloproteins is of importance to realize the relationship between metal ion and biological behavior, identification of metalloproteins is difficult. Here, we have developed three types of new polyacrylamide gel electrophoresis (PAGE) specific to metalloproteins and protein-bound metal ions in biological samples: metal ion contaminant sweeping (MICS)-Blue Native (BN)-PAGE; metal ion detection PAGE; and holo-apo conversion two dimensional MICS-PAGE.
In MICS-BN-PAGE, all metal contaminants were excluded from the gel, the separation buffer and the instruments, resulting in protein separation under metal-free conditions. In metal detection PAGE, metal ions (Fe, Cu and others) were separated and fluorescently detected with high sensitivity (to the ppt level) by the addition of probes. In holo-apo conversion 2D MICS-BN-PAGE, metalloproteins were exclusively isolated off of the diagonal line.

Free Research Field

分析化学