Isolation and characterisation of a 17-kDa staphylococcal heparin-binding protein with broad specificity

Abstract

A previous study reported the ability of staphylococci to bind heparin and heparindependent host growth factors. The present study isolated and identi®ed heparin-and basic ®broblast growth factor (bFGF)-binding surface components of S. epidermidis strain RP12 and S. haemolyticus strain SM 131. The staphylococcal heparin-binding component(s) were puri®ed by af®nity chromatography on heparin-Sepharose and a major heparin-binding protein, here designated HBP, was identi®ed by immunoblot in these two coagulase-negative staphylococcal (CNS) species. The HBP was shown to be acidic with an approximate pI of 4.6 and a molecular mass around 17 kDa. The binding of heparin to HBP was inhibited by heparin, fucoidan, pentosan polysulphate and various other sulphated polysaccharides, but not by non-sulphated compounds. However, the puri®ed HBP from both S. epidermidis and S. haemolyticus revealed broad speci®city, and also bound bFGF, thrombospondin, von Willebrand factor and, weakly, ®brinogen. The N-terminal sequences of the 17-kDa HBP from S. epidermidis and S. haemolyticus showed only limited identity. Comparison of the ®rst 15 amino acid residues derived from either strain with known sequences in the protein databases revealed no close similarities. Taken together, these results suggest that the adhesion of at least some CNS to host sulphated glycosaminoglycans may be mediated by a previously uncharacterised group of surface proteins.

Hyaluronan Modulation Impacts Staphylococcus aureus Biofilm Infection

“…This pathogen might contain surface proteins that bind HA and facilitate its incorporation into the biofilm structure. Heparin and eDNA binding proteins have been identified in a number of bacteria, including S. aureus (45)(46)(47)(48)(49)(50), lending support to this idea. Due to the high degree of HA association with the individual cell surfaces in S. aureus biofilm (Fig.…”

Heparin Stimulates Staphylococcus aureus Biofilm Formation

“…These data suggests that heparin stimulates the formation of adhesion molecules that make S. aureus better able to adhere to one another in either a heparindependent (where heparin acts as a cross-bridge) or heparinindependent manner. Consistent with this hypothesis, several groups have evidence suggesting that S. aureus codes for a heparin binding protein (14,16,37). Pascu and colleagues found that 30 of 38 coagulase-negative staphylococci were able to aggregate to heparin-coated beads and this binding was inhibited by several sulfated molecules (48).…”

Affinity-Based Methods for the Separation of Parasite Proteins

“…Although some parasites can bind to multiple GAGs (Coppi et al, 2007;Fallgren et al, 2001), heparan sulfate proteoglycan (HSPG) has been implicated in the recognition and/or invasion process of a wide range of pathogens, including viruses, bacteria and protozoan parasites (Bambino-Medeiros et al, 2011;Dalrymple & Mackow 2011;Yan et al, 2006;). Despite the role of heparin-binding proteins in many physiological and pathological processes, the basis of the heparin-protein interaction at the molecular level is still unclear.…”

“…HBPs from S. aureus have been demonstrated to be sensitive to heat and proteases, such as pronase E, proteinase K, pepsin and chymotrypsin (Liang et al, 1992). Interestingly, a 17-kDa heparin-binding protein with pI 4.6 has also been isolated from S. epidermis and S. haemolyticus, but the amino acid sequence similarity is low between these two organisms (MXTAHSYTXKYNGYTAN and MATQTKGYYYSYNGYV, respectively) and other bacterial HBPs (Fallgren et al, 2001). …”