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Split‐BioID: a proximity biotinylation assay for dimerization‐dependent protein interactions
Author(s) -
De Munter Sofie,
Görnemann Janina,
Derua Rita,
Lesage Bart,
Qian Junbin,
Heroes Ewald,
Waelkens Etienne,
Van Eynde Aleyde,
Beullens Monique,
Bollen Mathieu
Publication year - 2017
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.12548
Subject(s) - biotinylation , dna ligase , protein subunit , phosphatase , mutant , biotin , complementation , biochemistry , protein fragment complementation assay , enzyme , chemistry , microbiology and biotechnology , biology , gene
The biotin identification (BioID) protocol uses a mutant of the biotin ligase BirA (BirA*) fused to a protein‐of‐interest to biotinylate proximate proteins in intact cells. Here, we show that two inactive halves of BirA* separately fused to a catalytic and regulatory subunit of protein phosphatase PP1 reconstitute a functional BirA* enzyme upon heterodimerization of the phosphatase subunits. We also demonstrate that this BirA* fragment complementation approach, termed split‐BioID, can be used to screen for substrates and other protein interactors of PP1 holoenzymes. Split‐BioID is a novel and versatile tool for the identification of (transient) interactors of protein dimers.