The quantification of ratios between apo to holo types of metal binding protein : a new indicator of the oxidative stress in cells

我们应用纳米电喷雾电离质谱法（NanoESIMS）对载子型和全型金属蛋白（铜/锌结合超氧化物歧化酶：SOD-1、金属硫氨酸）进行定量分析，以阐明家族性肌萎缩性侧索硬化症等神经退行性疾病的病因。采用金属蛋白标准品和纯化金属蛋白，研究了超灵敏质谱法定量测定金属蛋白含量的最佳条件。1)考察了样品溶液的pH范围、有机溶剂的含量、加热毛细管的温度等条件。2)采用电感耦合等离子体质谱（ICPMS）代替纳米esims技术。采用商用SOD-1标准品，修改了分析条件，仍不能检测到二聚体、全息型。我们放弃了纳米esims技术最优条件的构建。另一方面，我们应用了ICPMS技术或SOD-1的全型转化，成功实现了金属蛋白的全型。该技术（ICPMS）是定量测定载脂蛋白与全型金属蛋白比率的有力工具。

We applied nano-electrospray ionization mass spectrometry (NanoESIMS) for the quantification of the rations between apo to holo types of metalloproteins (copper/zinc binding superoxide dismutase:SOD-1, metallothioneine) to clarify the etiology of neurodegenerative diseases such as familial amyotrophic lateral sclerosis. We examined the optimal conditions of ultra-sensitive mass spectrometric quantification of the ratios using metalloproteins standard and purified metalloproteins.1) We examined the condition of pH range of sample solutions, contents of organic solvent and temperature of heated capillary.2) We applied inductively coupled plasma mass spectrometry (ICPMS) instead of nanoESIMS technique.Using the commercial SOD-1 standard, we could not detect dimer, holo type even though we modified some analytical conditions. We give up the construction of the optimal condition of NanoESIMS technique. On the other hand, we applied the ICPMS technique or the de e turn of holo type of SOD-1 and succeeded holo type of the metalloprotem. This technique (ICPMS) was a powerful tool of the quantification of the ratios between apo to holo types of metalloproteins.