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 introduce 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 increase sample throughput and improve protein identification and quantification reproducibility. We applied this pipeline to delineate subcellular proteomes across various compartments. Using the 5HT2A serotonin receptor as a model, we studied temporal changes of proximal interaction networks induced by receptor activation. Additionally, we modified the p

SUBMITTER: Xiaofang Zhong 

PROVIDER: S-SCDT-10_1038-S44320-024-00049-2 | biostudies-other |

REPOSITORIES: biostudies-other