The purpose of this proteomic study was to identify RNA-binding proteins that specifically interact with the tRNA-derived fragment tRF-1432 in breast cancer cells. tRF-1432, a 5′-tRNA-derived stress-induced RNA generated from tRNA^Val-CAC-2-1, has been implicated in post-transcriptional regulation through its potential binding to RNA-binding proteins. To explore this mechanism, biotin-labeled tRF-1432 probes were used in RNA pulldown assays performed in MCF-7 and MDA-MB-231 breast cancer cell lines. The eluates were subjected to LC-MS/MS analysis, enabling the identification of candidate proteins potentially associated with tRF-1432.