PUBLICATION
In vivo proteomic mapping through GFP-directed proximity-dependent biotin labelling in zebrafish
- Authors
- Xiong, Z., Lo, H.P., McMahon, K.A., Martel, N., Jones, A., Hill, M.M., Parton, R.G., Hall, T.E.
- ID
- ZDB-PUB-210217-9
- Date
- 2021
- Source
- eLIFE 10: (Journal)
- Registered Authors
- Parton, Robert G.
- Keywords
- cell biology, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Biotin/pharmacology*
- Biotinylation
- Caveolins/metabolism
- Endothelial Cells/metabolism
- Green Fluorescent Proteins
- Membrane Proteins/metabolism
- Muscle, Skeletal/metabolism
- Nanoparticles
- Neurons/metabolism
- Protein Interaction Mapping
- Proteomics/methods*
- Staining and Labeling/methods*
- Zebrafish
- PubMed
- 33591275 Full text @ Elife
Citation
Xiong, Z., Lo, H.P., McMahon, K.A., Martel, N., Jones, A., Hill, M.M., Parton, R.G., Hall, T.E. (2021) In vivo proteomic mapping through GFP-directed proximity-dependent biotin labelling in zebrafish. eLIFE. 10:.
Abstract
Protein interaction networks are crucial for complex cellular processes. However, the elucidation of protein interactions occurring within highly specialised cells and tissues is challenging. Here we describe the development, and application, of a new method for proximity-dependent biotin labelling in whole zebrafish. Using a conditionally stabilised GFP-binding nanobody to target a biotin ligase to GFP-labelled proteins of interest, we show tissue-specific proteomic profiling using existing GFP-tagged transgenic zebrafish lines. We demonstrate the applicability of this approach, termed BLITZ (Biotin Labelling In Tagged Zebrafish), in diverse cell types such as neurons and vascular endothelial cells. We applied this methodology to identify interactors of caveolar coat protein, cavins, in skeletal muscle. Using this system, we defined specific interaction networks within in vivo muscle cells for the closely related but functionally distinct Cavin4 and Cavin1 proteins.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping