This project contains label-free quantitative proteomic datasets generated to compare the proteomes of wild-type Staphylococcus aureus USA300 and the ΔcrcB1ΔcrcB2 mutant. Three independent biological replicates were analyzed for each strain. Bacterial cultures were grown in TSB medium at 37 °C with shaking until the OD600 reached approximately 0.8. Cell pellets and corresponding culture supernatants were collected separately. Cell-associated proteins were extracted from bacterial pellets, whereas extracellular proteins were enriched from clarified and filtered culture supernatants by methanol precipitation. Proteins were reduced, alkylated, digested with trypsin, desalted, and analyzed by nanoLC-MS/MS using a Bruker timsTOF Pro 2 mass spectrometer operated in PASEF mode. Raw data were searched against the UniProt S. aureus USA300 TCH1516 protein database. Label-free quantification was performed separately for the cell-associated and culture-supernatant datasets. Differentially abundant proteins were identified using a fold-change cutoff of ≥ 1.2 or ≤ 0.83 and an FDR-adjusted P value < 0.05.