One-step purification of metallothionein extracted from two different sources |
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Authors: | Honda Rubens T Araújo Roziete Mendes Horta Bruno Brasil Val Adalberto L Demasi Marilene |
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Institution: | Instituto Nacional de Pesquisas da Amaz?nia, INPA, Manaus, AM, Brazil. |
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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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