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离子化质谱（NanoESIMS）的定量之间的比率apo holo类型的金属蛋白（铜/锌结合超氧化物歧化酶：SOD-1，金属硫蛋白），以澄清神经退行性疾病，如家族性肌萎缩侧索硬化症的病因。本研究以金属蛋白标准品和纯化的金属蛋白为研究对象，考察了超灵敏质谱定量的最佳条件：1）考察了样品溶液的pH范围、有机溶剂的含量和加热毛细管的温度; 2）采用电感耦合等离子体质谱（ICPMS）代替nanoESIMS技术，以商品SOD-1为标准品，检测不到二聚体，虽然我们修改了一些分析条件。我们放弃了NanoESIMS技术最佳条件的构建。另一方面，我们应用ICPMS技术或SOD-1的全息分型技术，成功地进行了金属蛋白的全息分型。该技术（ICPMS）是一个强有力的工具，定量的比例之间的apo和holo类型的金属蛋白。

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.