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Label Transfer Chemistry for the Characterization of Protein−Protein Interactions

2007; American Chemical Society; Volume: 129; Issue: 41 Linguagem: Inglês

10.1021/ja072904r

1943-2984

Бо Лю, Chase T. Archer, Lyle Burdine, Thomas G. Gillette, Thomas Kodadek,

Glycosylation and Glycoproteins Research

A new label transfer method is presented that overcomes most of the limitations of current systems. A protein of interest is tagged with a tetracysteine sequence (FlAsH receptor peptide (FRP)) that binds tightly and specifically to a chimeric molecule 3,4-dihydroxyphenylalanine-biotin-4',5'-bis(1,3,2-dithioarsolan-2-yl)fluorescein (DOPA-biotin-FlAsH). Upon brief periodate oxidation, the DOPA moiety is cross-linked to nearby surface-exposed nucleophiles. Boiling the products in excess dithiol dissolves the FlAsH-FRP interaction, resulting in transfer of the biotin tag to the partner proteins, allowing them to be identified by standard methods.