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Dev Biol 2022 Dec 01;492:133-138. doi: 10.1016/j.ydbio.2022.10.005.
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TurboID functions as an efficient biotin ligase for BioID applications in Xenopus embryos.

Kanzler CR , Donohue M , Dowdle ME , Sheets MD .

BioID is a proximity labeling strategy whose goal is to identify in vivo protein-protein interactions. The central components of this strategy are modified biotin ligase enzymes that promiscuously add biotin groups to proteins in close proximity. The transferred biotin group provides a powerful tag for purification and thus identification of interacting proteins. While a variety of modified biotin ligases were created for BioID, the original enzymes were inefficient, required long incubation times, and high intracellular biotin concentrations for protein labeling. These limitations hinder the application of BioID in contexts such as developing embryos where processes such as cell division and cell fate decisions occur rapidly. Recently, a new biotin ligase called TurboID was developed that addressed many of the deficiencies of previous enzymes. In this paper we compare TurboID to the BioID2 biotin ligase in developing Xenopus embryos. We find that the TurboID enzyme has several advantages over the BioID2 enzyme. TurboID labels proteins efficiently without the addition of additional biotin and occurs at a range of temperatures compatible with the culturing of Xenopus embryos. Biotinylation events occurred rapidly and were limited by TurboID expression and not its activity. Thus, TurboID is an efficient tool for BioID applications in Xenopus embryos and its use should facilitate the identification of interacting proteins in specific networks and complexes during Xenopus development.

PubMed ID: 36270327
PMC ID: PMC9643680
Article link: Dev Biol
Grant support: [+]

Species referenced: Xenopus laevis
Genes referenced: bicc1
GO keywords: enzyme activator activity [+]

Article Images: [+] show captions
References [+] :
Ahmed, Pre-hybridisation: an efficient way of suppressing endogenous biotin-binding activity inherent to biotin-streptavidin detection system. 2014, Pubmed