Toxicology Program, Utah State University, 84322, Logan, UT.
Biol Trace Elem Res. 1983 Jun;5(3):195-204. doi: 10.1007/BF02916623.
In vivo as well as in vitro protein-metal interaction was studied in cytosolic fractions from intestinal mucosal cells. Female Swiss-Webster mice wre pretreated with cadmium (25 ppm) or copper (100 ppm) in drinking water for 3 weeks. Treatment groups were divided into subgroups receiving Cd or Cd+Cu for an additional 6 weeks. In the in vitro study, mucosal cytosol obtained from pretreated animals was incubated with Cd-109 or Cd-109+Cu. Proteins were separated by gel filtration chromatography and metals determined by furnace AAS or gamma-spectrometry. Cadmium-induced synthesis of metallothionein-like proteins (MTP) in cytosol was indicated by increased Cd in those eluted fractions corresponding to the molecular weight of purified equine renal metallothionein. This cadmium level reached a plateau after 3 weeks of cadmium treatment. In addition, an increased amount of cadmium bound to MTP was noted when copper was added to cadmium in drinking water of mice pretreated with copper. This was not the case for Cd-pretreated animals. The in vitro experiments produced similar results, in that MTP fractions retained a greater percentage of Cd when animals were pretreated with copper compared to controls. Cadmium pretreatment resulted in even higher amounts of cadmium bound to MTP. The existence of a Cd as well as a separate Cu MTP, each with specific metal-binding properties, is suggested.
在肠黏膜细胞胞质部分进行了体内和体外的蛋白质-金属相互作用研究。雌性瑞士-韦伯斯特小鼠用饮用水中的镉(25ppm)或铜(100ppm)预处理 3 周。治疗组分为亚组,分别用 Cd 或 Cd+Cu 再处理 6 周。在体外研究中,用预处理动物的黏膜胞质液孵育 Cd-109 或 Cd-109+Cu。蛋白质通过凝胶过滤色谱分离,金属通过熔炉 AAS 或伽马光谱法测定。在胞质液中,Cd 诱导合成了金属硫蛋白样蛋白(MTP),表明与纯化的马肾金属硫蛋白分子量相对应的洗脱部分中的 Cd 增加。在镉处理 3 周后,镉水平达到了一个平台。此外,当在预先用铜处理的小鼠饮用水中添加铜时,与 Cd 结合的 MTP 的量也增加了。而对于 Cd 预处理的动物,则不是这样。体外实验产生了类似的结果,即当动物用铜预处理时,MTP 部分保留了更高比例的 Cd,与对照相比。镉预处理导致与 MTP 结合的镉量甚至更高。提示存在 Cd 和单独的 Cu-MTP,每种都具有特定的金属结合特性。