Zaia J, Fabris D, Wei D, Karpel R L, Fenselau C
Department of Chemistry and Biochemistry and the University of Maryland Cancer Center, University of Maryland Baltimore County, 21250, USA.
Protein Sci. 1998 Nov;7(11):2398-404. doi: 10.1002/pro.5560071117.
The capabilities of electrospray ionization mass spectrometry are demonstrated for monitoring the flux of metal ions out of and into the metalloprotein rabbit liver metallothionein and, in one example, chlorambucil-alkylated metallothionein. Metal ion transfers may be followed as the reactions proceed in situ to provide kinetic information. More uniquely to this technique, metal ion stoichiometries may be determined for reaction intermediates and products. Partners used in these studies include EDTA, carbonic anhydrase, a zinc-bound hexamer of insulin, and the core domain of bacteriophage T4 gene 32 protein, a binding protein for single-stranded DNA.
电喷雾电离质谱法的能力通过监测金属离子进出金属蛋白兔肝金属硫蛋白以及(在一个例子中)苯丁酸氮芥烷基化的金属硫蛋白的通量得以展现。随着反应在原位进行,可以跟踪金属离子的转移以提供动力学信息。这项技术更独特的是,可以确定反应中间体和产物的金属离子化学计量。这些研究中使用的伙伴包括乙二胺四乙酸(EDTA)、碳酸酐酶、锌结合的胰岛素六聚体以及噬菌体T4基因32蛋白的核心结构域,一种单链DNA结合蛋白。