Biotinylation protein is one technique used in biotechnology for tagging the proteins. In BioID, the protein of interest is fused to a “promiscuous” biotin ligase and stably expressed in cells; the fusion protein biotinylated proximal proteins only when excess biotin is added to the medium. High-affinity avidin/streptavidin-beads are used to capture biotinylated candidates in a …
Category Archives: Biotinylated Fusion Protein
How to identify biotinylated proteins? Short fast 1 minute answer!
Analogous to DamID, the biotin ligase is fused to a protein of interest, and then introduced into mammalian (or other) cells where it will biotinylate vicinal proteins upon supplementation of the culture medium with biotin. Biotinylated proteins can then be selectively isolated and identified by conventional methods, most notably mass spectrometry.
What is a promiscuous biotin ligase fusion protein? Short fast 1 minute answer!
A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells Kyle J. Roux,1,2Dae In Kim,1,2Manfred Raida,3and Brian Burke4