FIG. 4.

Ligand affinity blotting demonstrated binding of rSgrA to nidogen and fibrinogen. (A) Human fibrinogen and fibronectin were separated through SDS-PAGE, while nidogen 1 and nidogen 2 were separated through native PAGE followed by Coomassie blue staining (indicated by “C”). The ligand affinity blots were probed with rSgrA (fibrinogen; left part) or biotinylated rSgrA (nidogen 1 and nidogen 2) and are indicated by an “L.” (B) Reciprocal ligand affinity binding assays. rSgrA was separated through SDS-PAGE and stained with Coomassie (lane C). Anti-His monoclonal antibodies (L1) or biotinylated ligands including fibrinogen (L2), nidogen 1 (L3), nidogen 2 (L4), and a negative control (biotinylated fibronectin; L5) were allowed to bind to rSgrA and were detected using a streptavidin conjugate. (C) Ligand affinity blotting under reducing conditions demonstrated binding of rEcbA to collagen type V and fibrinogen and not to fibronectin.