In Staphylococcus aureus, the target of RNAIII activating protein (TRAP) is a membrane-associated protein whose C-terminus can be used as a vaccine to provide protection against staphylococcal infection. Here, we show for the first time by surface plasmon resonance and enzyme-linked immunosorbent assay that TRAP can specifically bind lysozyme and lysostaphin through its C-terminus (amino acids 155-167) and enhance lysozomal activities in vitro. It was also found that the traP mutant strain is more resistant to lysostaphin than wild-type. Our previous data showed that the C-terminus of TRAP might be extracellular. So our results suggested that the C-terminus of TRAP could act as the specific targeting protein of the lysozyme/lysostaphin on the S. aureus cell wall and the biological significance of the interaction might be to facilitate lysozyme/lysostaphin-mediated cell lysis.