1. Split-BioID: a proximity biotinylation assay for dimerization-dependent protein interactions.
- Author
-
De Munter, Sofie, Görnemann, Janina, Derua, Rita, Lesage, Bart, Qian, Junbin, Heroes, Ewald, Waelkens, Etienne, Van Eynde, Aleyde, Beullens, Monique, and Bollen, Mathieu
- Subjects
BIOTIN ,PROTEIN-ligand interactions ,PHOSPHOPROTEINS ,PROTEIN-protein interactions ,PHOSPHATASES - 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. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF