One-step purification of metallothionein extracted from two different sources
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 metallothione...
| Autor principal: | Honda, Rubens Tomio |
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| Outros Autores: | Araújo, Roziete Mendes, Horta, Bruno Brasil, Val, Adalberto Luis, Demasi, Marilene |
| Grau: | Artigo |
| Idioma: | English |
| Publicado em: |
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
2020
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| Acesso em linha: |
https://repositorio.inpa.gov.br/handle/1/18836 |
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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. © 2005 Elsevier B.V. All rights reserved. |