CLONING, SEQUENCING, AND EXPRESSION OF A FIBRONECTIN FIBRINOGEN-BINDING PROTEIN FROM GROUP-A STREPTOCOCCI/

Lipoteichoic acid and several streptococcal proteins have been reporte d to bind fibronectin (Fn) or fibrinogen (Fgn), which may serve as hos t receptors. We searched for such proteins by screening a library of g enes from M type 5 group A streptococci cloned into Escherichia coli. Lysates of clones were probed with biotinylated Fn and biotinylated Fg n. One clone expressed a 54-kDa protein that reacted with Fn and Fgn. The protein, termed FBP54, was purified and used to immunize rabbits. Anti-FBP54 serum reacted with purified, recombinant FBP54 and with a p rotein of similar electrophoretic mobility in extracts of M type 5, 6, and 24 streptococci. Anti-FBP54 serum also reacted with 5 of 15 strai ns of intact, live streptococci, suggesting that PBP54 may be a surfac e antigen. Southern blot analysis confirmed that the gene is found in group A streptococci but not in Staphylococcus aureus or E. coli. The cloned gene was sequenced and contained an open reading frame encoding a protein with a calculated molecular weight of 54,186. Partial amino acid sequencing of purified FBP54 confirmed that this open reading fr ame encoded the protein. As determined by utilizing fusion proteins co ntaining truncated forms of FBP54, the primary Fn/Fgn-binding domain a ppears to be contained in residues 1 to 89. These data suggest that FB P54 may be a surface protein of streptococci that reacts with both Fn and Fgn and therefore may participate in the adhesion of group A strep tococci to host cells.