One-step purification of metallothionein extracted from two different sources

被引：10

作者：

Honda, RT

Araújo, RM

Horta, BB

Val, AL

Demasi, M

机构：

[1] INPA, Manaus, Amazonas, Brazil

[2] Univ Sao Paulo, Inst Biociencias, Sao Paulo, Brazil

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2005年 / 820卷 / 02期

关键词：

metallothionein; environmental sciences; Ni and Cd binding; metal-chellating affinity chromatography;

D O I：

10.1016/j.jchromb.2005.03.017

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a one-step purification of hepatic metallothionein from the Amazon fish Colossoma macropomum injected with cadmium and from the copper-loaded metallothionein from the yeast Saccharomyces cerevisiae, performed by affinity chromatography through metal-chelating columns. Yeast metallothionein was purified from Cu2+-loaded resin and eluted by a continuous EDTA gradient whereas hepatic metallothionein extracted from fishes was purified by W, -loaded resin and eluted by a continuous imidazol gradient. Purified metallothioneins were evaluated by SDS-PAGE and characterized by UV spectra of the apo- and Cd2+-loaded protein. This method allowed high purity and yield as well as rapid one-step extraction of both metal-loaded and apoprotein. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：205 / 210

页数：6

共 22 条

[1] Accumulation and soluble binding of cadmium, copper, and zinc in the polychaete Hediste diversicolor from coastal sites with different trace metal Bioavailabilities [J].