Faculty, Staff and Student Publications
Publication Date
1-1-2025
Journal
Biotechniques
DOI
10.1080/07366205.2026.2619158
PMID
41607265
Abstract
Proximity labeling has become a powerful technique for mapping protein-protein interactions under physiologically relevant conditions, with TurboID offering high enzymatic activity and rapid labeling. However, its application in endothelial systems has been limited, partly due to the presence of endogenous biotin in specialized media, which reduces labeling specificity. Here, we optimized TurboID-mediated proximity labeling in brain microvascular endothelial cells by addressing two key challenges: endogenous biotin interference and cost-effective depletion. We discovered that endothelial cell medium contains substantial biotin levels, which saturate TurboID labeling and obscure the effects of exogenous biotin. Using High Capacity NeutrAvidin
Keywords
Biotin, Endothelial Cells, Biotinylation, Humans, Protein Interaction Mapping, Animals, Streptavidin, Brain, Staining and Labeling, Avidin, Cells, Cultured, Proximity labeling, TurboID, biotin depletion, endothelial cells, protein-protein interaction
Published Open-Access
yes
Recommended Citation
Jiang, Ying; Qu, Kuizhi; Dai, Mengjun; et al., "Optimization of Proximity Labeling in Endothelial Cells: Overcoming Endogenous Biotin Interference and Cost Barriers" (2025). Faculty, Staff and Student Publications. 6467.
https://digitalcommons.library.tmc.edu/uthgsbs_docs/6467
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