Kabziński A K
University of Lódź, Department of General and Inorganic Chemistry, Poland.
Biomed Chromatogr. 1998 Sep-Oct;12(5):281-90. doi: 10.1002/(SICI)1099-0801(199809/10)12:5<281::AID-BMC748>3.0.CO;2-8.
Intoxication with heavy metals results in numerous poisonings and diseases. They disturb metabolism of the system, are the source of cancer, degeneration changes and others. As a result of kidney damage the urine of people exposed to heavy metals contains different low molecular weight proteins, oligopeptides and amino acids, indicating pathological changes. One of the proteins is a very specific metallopolythiopolypeptide--metallothionein (MT). Based on earlier investigations, a very good correlations has been found between the contents of metallothionein in urine and plasma and the concentration of heavy metals in the blood, urine, kidneys, liver and brain and general in level of exposition to heavy metals. The aim of our investigations was to carry out quantitative isolation of Zn-thionein (Zn-Th), in order to determine the level of exposition to heavy metals. For Zn-Th protein isolation by covalent affinity chromatography with thiol-disulphide interchange (CAC-TDI) was applied, which is a modern technique of separation of a high affinity, good repeatability and reproducibility, allowing specific isolation of the thiol-proteins CAC-TDI gel was used as a solid-phase extraction (SPE) support for preconcentration of Zn-Th protein and Zn bonded with Zn-Th from water, rine, plasma and breast milk samples. The investigations showed unfavourable effect of the support on separation of thiol proteins and good correlation between the concentration of MTs protein added to water, plasma and urine and the concentration of protein indirectly determined via atomic absorption spectrometric (AAS) method, by preconcentration on SPE support metals formerly bound with MT protein and absorbed on CAC-TDI gel and calculated from metals concentration. The present paper is a continuation of earlier experiments on quantitation of Hg-thionein and Cd-thionein in physiological fluids and homogenates.
重金属中毒会导致大量中毒事件和疾病。它们扰乱系统的新陈代谢，是癌症、退行性病变及其他疾病的根源。由于肾脏受损，接触重金属的人的尿液中含有不同的低分子量蛋白质、寡肽和氨基酸，这表明存在病理变化。其中一种蛋白质是非常特殊的金属聚硫多肽——金属硫蛋白（MT）。基于早期研究，已发现尿液和血浆中金属硫蛋白的含量与血液、尿液、肾脏、肝脏和大脑中重金属的浓度以及总体重金属暴露水平之间存在非常良好的相关性。我们研究的目的是对锌硫蛋白（Zn-Th）进行定量分离，以确定重金属暴露水平。采用了通过硫醇 - 二硫化物交换的共价亲和色谱法（CAC-TDI）分离Zn-Th蛋白，这是一种具有高亲和力、良好重复性和再现性的现代分离技术，能够特异性分离硫醇蛋白。CAC-TDI凝胶用作固相萃取（SPE）支持物，用于从水、尿液、血浆和母乳样品中预富集Zn-Th蛋白以及与Zn-Th结合的锌。研究表明该支持物对硫醇蛋白分离有不利影响，并且添加到水、血浆和尿液中的MTs蛋白浓度与通过原子吸收光谱法（AAS）间接测定的蛋白浓度之间存在良好相关性，该间接测定是通过在SPE支持物上预富集先前与MT蛋白结合并吸附在CAC-TDI凝胶上的金属，然后根据金属浓度计算得出。本文是早期关于生理流体和匀浆中汞硫蛋白和镉硫蛋白定量实验的延续。
