Objective To identify Rab25-interacting proteins in gastric cancer cells using tandem affinity purification-mass spectrometry (TAP-MS).Methods An SGC7901 cell strain that could stably express Rab25 was constructed using a StrepII-6His-EGFP-Rab25 expressing plasmid by lentivirus transfection technology. After StrepII-6His tandem affinity purification, mass spectrometry was carried out to analyze the interaction protein complex of Rab25. To further verify their interactions, a protein-protein network was constructed and the authenticity of interactions was verified by co-immunoprecipitation in gastric cancer cells.Results A total of 26 Rab25 interacting proteins were identified by TAP-MS technique, which were enriched in the cancer process, apoptosis, NOTCH-mediated pathway and microtubule-based movement. Co-immunoprecipitation analysis confirmed that Itgb1 interacted with Rab25, and that they collaborated to promote the invasion of SGC7901 cells according to Transwell invasion assays.Conclusions This study helps to draw a detailed map of Rab25 interactions and provides clues for treatment of gastric cancer.
Key words
gastric cancer /
Rab25 /
protein-protein interaction /
tandem affinity purification /
mass spectrometry
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