A method for detecting protein targets of an enzyme using a biotin ligase

Abstract

A method of detecting an interaction or proximity between a fusion protein and a target protein. The method may include: a. providing a fusion protein and/or a recombinant nucleic acid molecule encoding said fusion protein, said fusion protein including an enzyme or enzyme complex member fused to a promiscuous biotin ligase, b. inhibiting the enzyme function of the fusion protein or the enzyme complex, c. biotinylating a target protein that interacts with and/or is in proximity to the fusion protein, d. enriching the biotinylated target protein, and identifying the isolated target protein using mass spectrometry (MS). The invention further relates to a fusion protein comprising a proteasome complex protein fused to a promiscuous biotin ligase, a cell line and a mouse expressing said fusion protein and a kit for performing the disclosed methods.

Dependencies are inferred from the claim text.