Pan A, Tie F, Ru B, Li L, Shen T
Department of Biology, Peking University, Beijing, China.
Biomed Chromatogr. 1992 Jul-Aug;6(4):205-11. doi: 10.1002/bmc.1130060411.
Zn-metallothioneins (MT-1 and MT-2) were isolated and purified from Wistar rat liver induced by subcutaneous injection with cadmium chloride over a short time. Instead of Sephadex G-50 and DEAE Sephadex A-50, new chromatographic media produced by Pharmacia, Sephacryl S-200, S-100 and DEAE Sepharose Fast Flow were used in the purification of metallothioneins. The time required for purification with the new method was only 1/3 that required with the usual method and had the same purification effect and rate of recovery. The number of mercapto groups measured with modified Ellman's reagent and cysteine as standard is 20 in MT molecules. Zn and Cd concentrations in each fraction were measured by single sweep polarography rather than atomic absorption spectrophotometry. MT-1 and MT-2 contained 6 gram atoms of zinc, but no cadmium. Purified MT-1 and MT-2 were shown by high performance liquid chromatographic analysis to be highly homogeneous and had an amino acid composition similar to that of Cd-MT.
通过短时间皮下注射氯化镉诱导的Wistar大鼠肝脏中分离并纯化出锌金属硫蛋白（MT-1和MT-2）。在金属硫蛋白的纯化过程中，使用了Pharmacia公司生产的新型色谱介质Sephacryl S-200、S-100和DEAE Sepharose Fast Flow，而不是Sephadex G-50和DEAE Sephadex A-50。用新方法纯化所需的时间仅为常规方法所需时间的1/3，且具有相同的纯化效果和回收率。以改良的Ellman试剂和半胱氨酸为标准测定，MT分子中的巯基数量为20个。各馏分中的锌和镉浓度通过单扫描极谱法而非原子吸收分光光度法测定。MT-1和MT-2含有6克原子的锌，但不含镉。高效液相色谱分析表明，纯化后的MT-1和MT-2高度均一，其氨基酸组成与镉金属硫蛋白相似。
