title: Large-scale identification of bacteria-host crosstalk by affinity chromatography: capturing the interactions of Streptococcus suis proteins with host cells.
abstract: Protein-protein interactions between bacteria and their hosts are responsible for all types of infection processes. The investigation of the bacteria-host crosstalk can provide a comprehensive understanding of the pathogenesis of bacterial disease. Despite scattered efforts in this field, a systematic identification of interactions between host and bacterial proteins remains unavailable. Here, we develop ACSP (affinity chromatography-based surface proteomics), which combines affinity chromatography and shotgun proteomics (LC-MS/MS), to investigate the interactions on a large-scale. Using ACSP, the potential surface interacting proteins (SIPs) of Streptococcus suis serotype 2 (SS2) were captured by the chromatographic resin, which was immobilized with the native surface molecules of Hep-2 cells. And then 40 potential SIPs were identified from the preys by LC-MS/MS, including 3 SIPs that have been previously reported in the literature. We selected 8 important SIPs and confirmed their ability to adhere to Hep-2 cells. Additionally, 3 newly identified SIPs, or their polyclonal antibodies, were found to significantly inhibit the adherence of SS2 to Hep-2 cells, indicating their essential role in the interaction between SS2 and Hep-2 cells. Using this example, we show that ACSP represents a new valuable tool for investigating the bacteria-host interactions.
authors.affiliation: National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Chen
authors.forename: Anding
authors.initials: A
authors.lastname: Zhang
authors.forename: Zhongmin
authors.initials: Z
authors.lastname: Xu
authors.forename: Ran
authors.initials: R
authors.lastname: Li
authors.forename: Huanchun
authors.initials: H
authors.lastname: Chen
authors.forename: Meilin
authors.initials: M
authors.lastname: Jin
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D019921:Biotinylation; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D054884:Host-Pathogen Interactions; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry; D007106:Immune Sera / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011994:Recombinant Proteins / Q000378:metabolism; D017011:Streptococcus suis / Q000378:metabolism / Q000502:physiology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D055767:Immobilized Proteins; D007106:Immune Sera; D011994:Recombinant Proteins; D010751:Phosphopyruvate Hydratase