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BioID system for identification of Txnip interacting proteins. Myc-BirA∗ was fused to the aminoterminus of Txnip (BirA∗-Txnip) to screen for protein interactions. BirA∗ catalyzes a two-step reaction: first, generation of reactive biotinyl-AMP from biotin and ATP, and second, the attachment of that biotinyl-AMP to a specific lysine on an interacting/proximal protein. Streptavidin beads are used to affinity-purify biotinylated proteins, which then are analyzed by mass spectrometry.
