Expression, purification and structural analysis of a fibrinogen receptor FbsA from Streptococcus agalactiae

Streptococcus agalactiae is a leading cause of bacterial sepsis and meningitis in neonates. FbsA, a fibrinogen receptor of S. agalactiae is highly repetitive protein with each repeat containing 16 amino acids. The protein sequence of FbsA shows no homology to any known fibrinogen binding protein from other bacterial species, making it a unique fibrinogen receptor. FbsA is cloned, expressed in E. coli and purified. The recombinant protein shows a laddering pattern in SDS-PAGE gel because of its poor stability in solution. The instability of the protein is probably because of the presence Gln-Gly dipeptide in each repeat. The circular dichroism study of FbsA has shown that the protein is composed of alpha helices predominantly and random coils to a lesser extent, which agrees with the predicted secondary structure. Ab initio modeling of a single repeat shows that FbsA is made up of mainly alpha helix and the structural model of multiple repeats (3 or 4) suggests that the protein might adopt some form of a repeating helical structure and the overall conformation of the molecule might change depending on the number of repeats.