Characterization of arsenic-binding proteins and its application to health effect evaluation.

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We prepared arsenic-bound sepharoses and investigated arsenic-binding proteins by chromatography using a column packed with the arsenic-bound sepharoses. SDS-PAGE analyses showed no proteins bound to pentavalent arsenic specifically, while several proteins that have specific and high affinity for trivalent arsenic. We prepared promyelocytic leukemia (PML) expressing cells and applied the cytosolic proteins to the arsenic-binding column and the column was elutes with glutathione-containing solution. PML appeared to bind trivalent arsenic by forming ternary complex at RING finger domain of this protein. Western blot and immunofluorescent microscopic analyses suggested that PML were conjugation with small ubiquitin-like modifier proteins (SUMOylation) in response to arsenic exposure. The arsenic-bound sepharose column enabled us to identify proteins which have a high affinity to arsenic using this method.