Mann K G, Lawler C M, Vehar G A, Church W R
J Biol Chem. 1984 Nov 10;259(21):12949-51.
Preparations of bovine and human coagulation Factor V were analyzed for copper using both atomic absorption and atomic emission spectroscopy. All preparations of the bovine and human protein were found to contain copper ion at a ratio of 1 copper ion bound per mol (Mr = 330,000) of Factor V. As a result of copper binding and sequence homology between ceruloplasmin and Factor V, bovine Factor V and thrombin-activated Factor V (Va) were assessed with respect to their visible and near ultraviolet absorption spectra and to their ability to oxidize N,N-dimethyl-p-phenylenediamine (a substrate for ceruloplasmin). Factor V and Factor Va exhibited absorption spectra with no maxima at either 310 or 610 nm, indicating that the copper is not bound in a site analogous to Type I or Type III copper sites in ceruloplasmin. Further, Factor V and Factor Va are not capable of serving as catalysts for the oxidation of N,N-dimethyl-p-phenylenediamine under solution conditions that are optimum for ceruloplasmin oxidase activity. These data suggest that the copper ion bound to Factor V may be functionally and structurally distinct from the Type I and Type III copper ion bound to ceruloplasmin.
使用原子吸收光谱法和原子发射光谱法对牛和人凝血因子V制剂中的铜进行了分析。发现牛和人蛋白质的所有制剂均含有铜离子，其比例为每摩尔（Mr = 330,000）因子V结合1个铜离子。由于铜结合以及铜蓝蛋白与因子V之间的序列同源性，对牛因子V和凝血酶激活的因子V（Va）的可见光谱和近紫外吸收光谱以及它们氧化N，N-二甲基对苯二胺（铜蓝蛋白的一种底物）的能力进行了评估。因子V和因子Va的吸收光谱在310或610 nm处均无最大值，这表明铜不是以类似于铜蓝蛋白中I型或III型铜位点的方式结合。此外，在对铜蓝蛋白氧化酶活性最适宜的溶液条件下，因子V和因子Va不能作为氧化N，N-二甲基对苯二胺的催化剂。这些数据表明，与因子V结合的铜离子在功能和结构上可能与与铜蓝蛋白结合的I型和III型铜离子不同。
