A proximity proteomics pipeline with improved reproducibility and throughput.

ABSTRACT: Proximity labeling (PL) via biotinylation coupled with mass spectrometry (MS) captures spatial proteomes in cells. Large-scale processing requires a workflow minimizing hands-on time and enhancing quantitative reproducibility. We introduced a scalable PL pipeline integrating automated enrichment of biotinylated proteins in a 96-well plate format. Combining this with optimized quantitative MS based on data-independent acquisition (DIA), we increased sample throughput and improved protein identification and quantification reproducibility. We applied this pipeline to delineate subcellular proteomes across various compartments. Using the 5HT_2A serotonin receptor as a model, we studied temporal changes of proximal interaction networks induced by receptor activation. In addition, we m

SUBMITTER: Zhong X 

PROVIDER: S-EPMC11297269 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature