Molecular Mechanism by Which Surface Antigen HP0197 Mediates Host Cell Attachment in the Pathogenic Bacteria Streptococcus suis

Abstract

Streptococcus suis, one of the most important and prevalent pathogens in swine, presents a major challenge to global public health. HP0197 is an S. suis surface antigen that was previously identified by immunoproteomics and can bind to the host cell surface. Here, we investigated the interaction between HP0197 and the host cell surface glycosaminoglycans (GAGs) using indirect immunofluorescence and cell adhesion inhibition assays. In addition, we determined that a novel 18-kDa domain in the N-terminal region of HP0197 functions as the GAG-binding domain. We then solved the three-dimensional structures of the N-terminal 18-kDa and C-terminal G5 domains using x-ray crystallography. Based on this structural information, the GAG-binding sites in HP0197 were predicted and subsequently verified using site-directed mutagenesis and indirect immunofluorescence. The results indicate that the positively charged residues on the exposed surface of the 18-kDa domain, which are primarily lysines, likely play a critical role in the HP0197-heparin interaction that mediates bacterium-host cell adhesion. Understanding this molecular mechanism may provide a basis for the development of effective drugs and therapeutic strategies for treating streptococcal infections.Background: The function of the Streptococcus suis antigen HP0197 is unknown, but it may be potentially involved in pathogenesis.Results: A novel GAG-binding domain on HP0197 was identified, and its structure was determined.Conclusion: Three clusters of basic residues on an 18-kDa domain of HP0197 are critical for heparin binding and cell adhesion.Significance: Understanding this mechanism will contribute to the design of drugs to treat streptococcal infections. Streptococcus suis, one of the most important and prevalent pathogens in swine, presents a major challenge to global public health. HP0197 is an S. suis surface antigen that was previously identified by immunoproteomics and can bind to the host cell surface. Here, we investigated the interaction between HP0197 and the host cell surface glycosaminoglycans (GAGs) using indirect immunofluorescence and cell adhesion inhibition assays. In addition, we determined that a novel 18-kDa domain in the N-terminal region of HP0197 functions as the GAG-binding domain. We then solved the three-dimensional structures of the N-terminal 18-kDa and C-terminal G5 domains using x-ray crystallography. Based on this structural information, the GAG-binding sites in HP0197 were predicted and subsequently verified using site-directed mutagenesis and indirect immunofluorescence. The results indicate that the positively charged residues on the exposed surface of the 18-kDa domain, which are primarily lysines, likely play a critical role in the HP0197-heparin interaction that mediates bacterium-host cell adhesion. Understanding this molecular mechanism may provide a basis for the development of effective drugs and therapeutic strategies for treating streptococcal infections. Background: The function of the Streptococcus suis antigen HP0197 is unknown, but it may be potentially involved in pathogenesis. Results: A novel GAG-binding domain on HP0197 was identified, and its structure was determined. Conclusion: Three clusters of basic residues on an 18-kDa domain of HP0197 are critical for heparin binding and cell adhesion. Significance: Understanding this mechanism will contribute to the design of drugs to treat streptococcal infections.