The collagen-binding protein Cna is a prototype cell surface protein from Staphylococcus aureus which fulfils important physiological functions during pathogenesis. While it is established that Cna binds to collagen (Cn) via the high-affinity collagen hug mechanism, whether this protein is engaged in other ligand-binding mechanisms is poorly understood. Here, we use atomic force microscopy to demonstrate that Cna mediates attachment to two structurally and functionally different host proteins, i.e., the complement system protein C1q and the extracellular matrix protein laminin (Lam), through binding mechanisms that differ from the collagen hug. We show that single Cna-C1q and Cna-Lam bonds are much weaker than the high-affinity Cna-Cn bond and that their formation does not require the B-region of Cna. At the whole cell level, we find that bacterial adhesion to C1q-substrates involves only one (or two) molecular bond(s), while adhesion to Lam is mediated by multiple bonds, thus suggesting that multivalent or cooperative interactions may enhance the strength of adhesion. Both C1q and Lam interactions can be efficiently blocked by monoclonal antibodies directed against the minimal Cn-binding domain of Cna. These results show that Cna is a multifunctional protein capable of binding to multiple host ligands through mechanisms that differ from the classical collagen hug.
Keywords: C1q; Cna; Staphylococcus aureus; atomic force microscopy; binding mechanisms; force spectroscopy; laminin.