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毛细管电泳分析金属硫蛋白。

Analysis of metallothionein by capillary electrophoresis.

机构信息

Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czech Republic.

出版信息

J Chromatogr A. 2012 Feb 24;1226:31-42. doi: 10.1016/j.chroma.2011.10.015. Epub 2011 Oct 12.

DOI:10.1016/j.chroma.2011.10.015
PMID:22036087
Abstract

Metallothioneins (MTs) belong to proteins playing a key role in metal ion homeostasis, maintenance of the redox pool and free radical scavenging in both prokaryotic and eukaryotic cells. Strong interactions of the MTs with essential and non-essential metal ions as well as unique MT structure and behavior under various conditions are subjects of numerous studies. Among other analytical techniques, capillary electrophoresis (CE) has been proven to be an effective tool not only for determination of MT in biological samples, but also for the identification of its isoforms and sub-isoforms in various types of samples. Moreover, CE has a great potential to investigate MT-metal and MT-protein interactions, which has not been fully utilized yet. Thus, it is not surprising that numerous studies devoted to the optimization of CE conditions such as background electrolyte composition, electrolyte modifiers and/or capillary surface modifications have been carried out since MT's discovery in 1957. From the MTs' detection point of view, optical detectors including absorbance, laser-induced fluorescence have been employed. Also mass spectrometric detection coupled to the various ionization techniques including inductively coupled plasma (ICP) and electrospray ionization (ESI) has been utilized for detail MT characterization and sensitive determination. In this paper, articles published from eighties to 2011 are reviewed, presenting both optimization of key parameters of CE method for MT determination as well as utilization of CE as a routine analytical technique for further investigation of complex biological and biochemical processes where MT is a key component.

摘要

金属硫蛋白(MTs)属于在原核和真核细胞中在金属离子稳态、氧化还原池维持和自由基清除中起关键作用的蛋白质。MTs 与必需和非必需金属离子的强烈相互作用以及 MT 在各种条件下的独特结构和行为是众多研究的主题。在其他分析技术中,毛细管电泳(CE)已被证明不仅是测定生物样品中 MT 的有效工具,而且是鉴定各种类型样品中其同工型和亚型的有效工具。此外,CE 具有研究 MT-金属和 MT-蛋白相互作用的巨大潜力,但尚未得到充分利用。因此,自 1957 年发现 MT 以来,已经进行了大量致力于优化 CE 条件(如背景电解质组成、电解质修饰剂和/或毛细管表面修饰)的研究,这并不奇怪。从 MTs 的检测角度来看,已经采用了包括吸光度、激光诱导荧光在内的光学检测器。还利用与各种离子化技术(包括电感耦合等离子体(ICP)和电喷雾电离(ESI))相结合的质谱检测,对 MT 进行详细的特征描述和灵敏的测定。本文综述了从 80 年代到 2011 年发表的文章,介绍了 CE 方法用于 MT 测定的关键参数的优化,以及 CE 作为常规分析技术在进一步研究 MT 作为关键成分的复杂生物和生化过程中的应用。

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