Evaluation of different purification procedures for the electrochemical quantification of mussel metallothioneins
Evaluation of different purification procedures for the electrochemical quantification of mussel metallothioneins
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DOI：
10.1016/s0039-9140(02)00239-4
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发表时间：
2002-07-19
期刊：
影响因子：
6.1
通讯作者：
Pavicic, J
中科院分区：
文献类型：
--
作者：
Erk, M;Ivankovic, D;Pavicic, J
几种不同的程序应用于纯化金属硫蛋白（NIT）分离的贻贝（贻贝galloprovincialis）暴露于镉的消化腺：热处理（在70和85 degreesC），溶剂沉淀，和凝胶过滤。采用电化学方法测定MT含量。根据获得的结果，对每种手术进行了统计学评价。事后Scheffe检验的应用表明，所有应用的处理之间存在显著差异。热处理和溶剂沉淀可有效去除样品中的高分子量（HMW）蛋白质。在未处理以及热或溶剂处理的样品中，MT10同种型保持不变。MT20亚型通过热处理显著减少，并通过溶剂沉淀急剧减少。由于MT 20被认为是金属效应和暴露的“目标”NIT亚型，因此建议使用对MT 20组分影响较小的纯化程序。根据给出的结果，这是热处理。(C)2002 Elsevier Science B.V.保留所有权利。
Several different procedures were applied to purify metallothionein (NIT) isolated from the digestive glands of the mussels (Mytilus galloprovincialis) exposed to cadmium: heat treatment (at 70 and 85 degreesC), solvent precipitation, and gel-filtration. Using electrochemical method, the MT contents were determined. Based on the obtained results each procedure was statistically evaluated. Application of the post-hoc Scheffe test showed that there were significant differences between all the treatments applied. Heat treatment and solvent precipitation effectively remove high molecular weight (HMW) proteins from the samples. In untreated as well as in heat or solvent treated samples MT10 isoform remains unchanged. MT20 isoform is significantly reduced by heat treatment and drastically reduced by solvent precipitation. Due to the fact that MT20 is considered as 'target' NIT isoform for metal effect and exposure it is recommended to use the purification procedure which less affects MT20 fraction. According to the presented results it is the heat treatment. (C) 2002 Elsevier Science B.V. All rights reserved.