Adhesive surface proteins of Erysipelothrix rhusiopathiae bind to polystyrene, fibronectin, and type I and IV collagens

Erysipelothrix rhusiopathiaeis a gram-positive bacterium that causes erysipelas in animals and erysipeloid in humans. We found two adhesive surface proteins ofE. rhusiopathiaeand determined the nucleotide sequences of the genes, which were colocalized and designatedrspAandrspB. The two genes were present in all of the serovars ofE. rhusiopathiaestrains examined. The deduced RspA and RspB proteins contain the C-terminal anchoring motif, LPXTG, which is preceded by repeats of consensus amino acid sequences. The consensus sequences are composed of 78 to 92 amino acids and repeat 16 and 3 times in RspA and RspB, respectively. Adhesive surface proteins of other gram-positive bacteria, includingListeria monocytogenesadhesin-like protein,Streptococcus pyogenesprotein F2 and F2-like protein,Streptococcus dysgalactiaeFnBB, andStaphylococcus aureusCna, share the same consensus repeats. Furthermore, the N-terminal regions of RspA and RspB showed characteristics of the collagen-binding domain that was described for Cna. RspA and RspB were expressed inEscherichia colias histidine-tagged fusion proteins and purified. The recombinant proteins showed a high degree of capacity to bind to polystyrene and inhibited the binding ofE. rhusiopathiaeonto the abiotic surface in a dose dependent manner. In a solid-phase binding assay, both of the recombinant proteins bound to fibronectin, type I and IV collagens, indicating broad spectrum of their binding ability. It was suggested that both RspA and RspB were exposed on the cell surface ofE. rhusiopathiae, as were the bacterial cells agglutinated by the anti-RspA immunoglobulin G (IgG) and anti-RspB IgG. RspA and RspB were present both in surface-antigen extracts and the culture supernatants ofE. rhusiopathiaeFujisawa-SmR (serovar 1a) and SE-9 (serovar 2). The recombinant RspA, but not RspB, elicited protection in mice against experimental challenge. These results suggest that RspA and RspB participate in initiation of biofilm formation through their binding abilities to abiotic and biotic surfaces.