Separation and characterization of rat kidney isometallothioneins induced by exposure to inorganic mercury.

M A Morcillo, J Santamaría
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引用次数: 10

Abstract

High-performance liquid chromatography (HPLC) was applied to the separation of metallothionein (MT) isoforms from different tissues from a variety of eukaryotic species. Recently we reported an analytical method for 203Hg-metallothionein, which detects the radioisotope bound to each iso-MT after separation by HPLC on a size-exclusion column coupled with on-line radioactivity flow detection. The MTs can be separated as distinct isoprotein peaks by elution with alkaline buffer solution owing to cation-exchange chromatographic action. In the present work, renal MT from rats exposed to inorganic mercury was separated into four peaks by UV and 203Hg detection. Moreover, it was resolved into four components by non-denaturing polyacrylamide gel electrophoresis. The two major components correspond to MT-1 and MT-2, which were characterized by amino acid analysis. Finally, Hg induces and binds to both iso-MTs.

无机汞诱导大鼠肾异金属硫蛋白的分离与表征。
应用高效液相色谱法(HPLC)分离真核生物不同组织的金属硫蛋白(MT)异构体。最近,我们报道了一种分析203hg -金属硫蛋白的方法,该方法通过HPLC - scale -exclusion柱耦合在线放射性流检测,检测分离后与每个iso-MT结合的放射性同位素。由于阳离子交换层析的作用,用碱性缓冲溶液洗脱后,MTs可以被分离成不同的同工蛋白峰。本研究通过紫外和203Hg检测,将无机汞暴露大鼠肾MT分离为4个峰。并通过非变性聚丙烯酰胺凝胶电泳将其分解为4个组分。两个主要成分对应MT-1和MT-2,通过氨基酸分析表征。最后,汞诱导并结合到这两种异构体上。
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