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In Vivo Application of TurboID-based Proximity Labeling in Drosophila melanogaster
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TurboID functions as an efficient biotin ligase for BioID applications in Xenopus embryos
Charlotte R Kanzler1, Michael Donohue2, Megan E Dowdle2
1Department of Biomolecular Chemistry, School of Medicine and Public Health, University of, Wisconsin, Madison, United States; Cell and Molecular Biology Program, United States.
Developmental Biology
|October 21, 2022
Summary
TurboID is a highly efficient proximity labeling enzyme for identifying protein interactions in Xenopus embryos. It outperforms BioID2 by enabling rapid labeling without added biotin, crucial for studying fast developmental processes.
Area of Science:
- Molecular Biology
- Developmental Biology
- Biochemistry
Background:
- BioID (proximity-dependent biotinylation) identifies in vivo protein-protein interactions using modified biotin ligases.
- Original BioID enzymes had limitations: inefficiency, long incubation times, and high biotin requirements, hindering rapid developmental studies.
- TurboID was developed to overcome these BioID deficiencies.
Purpose of the Study:
- To compare the performance of TurboID and BioID2 proximity labeling enzymes in developing Xenopus embryos.
- To evaluate TurboID's efficiency and suitability for studying rapid biological processes in vivo.
Main Methods:
- Comparative analysis of TurboID and BioID2 in Xenopus embryos.
- Assessment of protein labeling efficiency, incubation time, and temperature compatibility.
- Evaluation of biotin supplementation requirements and labeling kinetics.
Main Results:
- TurboID demonstrates superior protein labeling efficiency compared to BioID2 in Xenopus embryos.
- TurboID achieves rapid biotinylation without requiring exogenous biotin supplementation.
- Labeling speed with TurboID is primarily limited by enzyme expression levels, not its catalytic activity.
Conclusions:
- TurboID is an efficient and versatile tool for proximity labeling applications in Xenopus embryos.
- Its rapid labeling kinetics and temperature compatibility make it ideal for studying dynamic developmental processes.
- TurboID facilitates the identification of protein interaction networks during Xenopus development.
