| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 9753326 | Journal of Chromatography B | 2005 | 6 Pages |
Abstract
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 Ni2+-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.
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Authors
Rubens T. Honda, Roziete Mendes Araújo, Bruno Brasil Horta, Adalberto L. Val, Marilene Demasi,