title: Opacity factor activity and epithelial cell binding by the serum opacity factor protein of Streptococcus pyogenes are functionally discrete.
abstract: Serum opacity factor (SOF) is a unique multifunctional virulence determinant expressed at the surface of Streptococcus pyogenes and has been shown to elicit protective immunity against GAS infection in a murine challenge model. SOF consists of two distinct domains with different binding capacities: an N-terminal domain that binds apolipoprotein AI and a C-terminal repeat domain that binds fibronectin and fibrinogen. The capacity of SOF to opacify serum by disrupting the structure of high density lipoproteins may preclude its use as a vaccine antigen in humans. This study generated mutant forms of recombinant SOF with reduced (100-fold) or abrogated opacity factor (OF) activity, for use as vaccine antigens. However, alterations introduced into the N-terminal SOF peptide (SOFDeltaFn) by mutagenesis to abrogate OF activity, abolish the capacity of SOF to protect against lethal systemic S. pyogenes challenge in a murine model. Mutant forms of purified SOFDeltaFn peptide were also used to assess the contribution of OF activity to the pathogenic processes of cell adhesion and cell invasion. Using latex beads coated with full-length SOF, SOFDeltaFn peptide, or a peptide encompassing the C-terminal repeats (FnBD), we demonstrate that adhesion to HEp-2 cells is mediated by both SOFDeltaFn and FnBD. The HEp-2 cell binding displayed by the N-terminal SOFDeltaFn peptide is independent of OF activity. We demonstrate that while the N terminus of SOF does not directly mediate intracellular uptake by epithelial cells, this domain enhances epithelial cell uptake mediated by full-length SOF, in comparison to the FnBD alone.
authors.affiliation: School of Biological Sciences, University of Wollongong, New South Wales 2522, Australia.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Gillen
authors.forename: Harry S
authors.initials: HS
authors.lastname: Courtney
authors.forename: Kai
authors.initials: K
authors.lastname: Schulze
authors.forename: Manfred
authors.initials: M
authors.lastname: Rohde
authors.forename: Mark R
authors.initials: MR
authors.lastname: Wilson
authors.forename: Anjuli M
authors.initials: AM
authors.lastname: Timmer
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Guzman
authors.forename: Victor
authors.initials: V
authors.lastname: Nizet
authors.forename: G S
authors.initials: GS
authors.lastname: Chhatwal
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Walker
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D016632:Apolipoprotein A-I / Q000235:genetics / Q000276:immunology; D001422:Bacterial Adhesion / Q000235:genetics / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000276:immunology* / Q000382:microbiology; D005340:Fibrinogen / Q000235:genetics / Q000276:immunology; D005353:Fibronectins / Q000235:genetics / Q000276:immunology; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D010447:Peptide Hydrolases / Q000235:genetics / Q000276:immunology*; D010455:Peptides / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D017434:Protein Structure, Tertiary / Q000235:genetics; D013290:Streptococcal Infections / Q000235:genetics / Q000276:immunology* / Q000517:prevention & control; D022541:Streptococcal Vaccines / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D013297:Streptococcus pyogenes / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity
chemical_list: D000942:Antigens, Bacterial; D016632:Apolipoprotein A-I; D001426:Bacterial Proteins; D005353:Fibronectins; D010455:Peptides; D022541:Streptococcal Vaccines; C021908:opacity factor; D005340:Fibrinogen; D010447:Peptide Hydrolases