Workflow enhancement of TurboID-mediated proximity labeling for SPY signaling network mapping.

ABSTRACT: TurboID-based proximity labeling coupled to mass spectrometry (PL-MS) has emerged as a powerful tool for mapping protein-protein interactions in both plant and animal systems. Despite advances in sensitivity, PL-MS studies can still suffer from false negatives, especially when dealing with low abundance bait proteins and their transient interactors. Protein-level enrichment for biotinylated proteins is well developed and popular, but direct detection of biotinylated proteins by peptide-level enrichment and the difference in results between direct and indirect detection remain underexplored. To address this gap, we compared and improved enrichment and data analysis methods using TurboID fused to SPY, a low-abundance O-fucose transferase, using an AAL-enriched SPY target library for cross-re

SUBMITTER: Grismer TS 

PROVIDER: S-EPMC10888891 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature