Journal
ELIFE
Volume 10, Issue -, Pages -Publisher
eLIFE SCIENCES PUBL LTD
DOI: 10.7554/eLife.64631
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Funding
- National Health and Medical Research Council [569542, 1045092, APP1044041, APP1099251]
- Australian Research Council [DP200102559, CE140100036]
- Australian Research Council [DP200102559] Funding Source: Australian Research Council
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This method utilizes proximity-dependent biotin labelling in zebrafish to conduct tissue-specific proteomic profiling, demonstrating its applicability in diverse cell types. Using this system, specific interaction networks within muscle cells for closely related Cavin4 and Cavin1 proteins were successfully defined.
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.
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