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 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.
Honda
Rubens T
RT
Instituto Nacional de Pesquisas da Amazônia, INPA, Manaus, AM, Brazil.
Araújo
Roziete Mendes
RM
Horta
Bruno Brasil
BB
Val
Adalberto L
AL
Demasi
Marilene
M
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005399:Fishes; D008668:Metallothionein / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D008668:Metallothionein