title: Split-BioID: a proximity biotinylation assay for dimerization-dependent protein interactions.
abstract: 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.
authors.affiliation: Laboratory of Biosignaling & Therapeutics, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.
authors.forename: Sofie
authors.initials: S
authors.lastname: De Munter
authors.affiliation: Laboratory of Biosignaling & Therapeutics, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.
authors.forename: Janina
authors.initials: J
authors.lastname: Görnemann
authors.affiliation: Protein Phosphorylation & Proteomics Lab, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.|SyBioMa, KU Leuven, Belgium.
authors.forename: Rita
authors.initials: R
authors.lastname: Derua
authors.affiliation: Laboratory of Biosignaling & Therapeutics, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.
authors.forename: Bart
authors.initials: B
authors.lastname: Lesage
authors.affiliation: Laboratory of Biosignaling & Therapeutics, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.
authors.forename: Junbin
authors.initials: J
authors.lastname: Qian
authors.affiliation: Laboratory of Biosignaling & Therapeutics, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.
authors.forename: Ewald
authors.initials: E
authors.lastname: Heroes
authors.affiliation: Protein Phosphorylation & Proteomics Lab, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.|SyBioMa, KU Leuven, Belgium.
authors.forename: Etienne
authors.initials: E
authors.lastname: Waelkens
authors.affiliation: Laboratory of Biosignaling & Therapeutics, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.
authors.forename: Aleyde
authors.initials: A
authors.lastname: Van Eynde
authors.affiliation: Laboratory of Biosignaling & Therapeutics, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.
authors.forename: Monique
authors.initials: M
authors.lastname: Beullens
authors.affiliation: Laboratory of Biosignaling & Therapeutics, KU Leuven Department of Cellular and Molecular Medicine, University of Leuven, Belgium.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Bollen
mesh_terms: D001681:Biological Assay / Q000379:methods*; D019921:Biotinylation; D019281:Dimerization; D005816:Genetic Complementation Test; D057809:HEK293 Cells; D006801:Humans; D025941:Protein Interaction Mapping; D015203:Reproducibility of Results
keywords: biotinylation; phosphatase-substrate mapping; protein-ligand screening; proximity labeling; reporter-fragment complementation