Characterization of Group B Streptococcal Glyceraldehyde-3-Phosphate Dehydrogenase: Surface Localization, Enzymatic Activity, and Protein-Protein Interactions
ABSTRACT During characterization of the surface antigens of serotype III group B streptococci (GBS), a protein with an apparent M(r) of approximately 173,500 migrating on a SDS--polyacrylamide gel was found to have an N-terminal amino acid sequence identical to that of the plasmin receptor (Plr) of group A streptococci, a surface-localized glyceraldehyde-3-phosphate dehydrogenase (GAPDH). This work begins to characterize GBS GAPDH and to assess its functional activity on the cell surface. The 1.0-kb gapC gene of GBS was amplified by PCR. plr and gapC demonstrated 87% homology. An anti-Plr monoclonal antibody reacted with GBS whole cells, suggesting GBS GAPDH is surface localized. Multiple serotypes of GBS demonstrated functional GAPDH on their surfaces. The anti-Plr monoclonal antibody recognized GBS protein bands of approximately 41 and 173.5 kDa, by Western blot. Presumably, these represent monomeric and tetrameric forms of the GAPDH molecule. GBS GAPDH was demonstrated by Western blot analysis to interact with lys- and glu-plasminogens. Fluid-phase GBS GAPDH interacted, by means of ELISA, with immobilized lys-plasminogen, glu-plasminogen, actin, and fibrinogen. Enzymatically active GAPDH, capable of binding cytoskeletal and extracellular matrix proteins, is expressed on the surface of GBS.
- SourceAvailable from: Shunqing Liang
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- "and Fischetti 1992; Seifert et al. 2003; Bergmann et al. 2004; Matta et al. 2010 "
ABSTRACT: Subcellullar localizations and cross-immunities of GAPDHs from six common pathogenic bacteria in aquaculture were investigated. Subcellullar localizations of GAPDHs of Edwardsiella tarda EIB202, Edwardsiella ictaluri ATCC33202, Aeromonas hydrophila LSA34, Vibrio anguillarum MVM425, Vibrio alginolyticus EPGS020401 and Vibrio harveyi VIB647 were analysed with Western blotting, indirect immunofluorescence and flow cytometry examinations. Immunoprotections of different recombinant GAPDHs against these pathogens were investigated with zebrafish model. Western blotting of subcellular extractions showed that all GAPDHs were secreted into extracellular medium and periplasmic space. In addition, GAPDHs were demonstrated to distribute in the outer membranes except MVM425 and VIB647. And, GAPDHs were confirmed to be present on the surface of these bacteria with indirect immunofluorescence and flow cytometry examinations. The remarkable cross-protective immunities of these recombinant GAPDHs were induced in zebrafish, and the relative protective survivals were almost over 60%. Localizations of GAPDHs from these pathogenic bacteria were similar to many other causative agents. And, GAPDHs could be important protective antigens and give remarkable cross-immunity against different pathogens. Recombinant GAPDH could be designed as a broad spectrum vaccine candidate against multiple microbial infections in aquaculture.Letters in Applied Microbiology 01/2012; 54(1):1-9. DOI:10.1111/j.1472-765X.2011.03164.x · 1.75 Impact Factor
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- "The GBS GAPDH is a moonlighting protein which, when present at the cell surface, interacts with cytoskeleton and extracellular matrix proteins and Plg . This bacterium also expresses a Fg receptor named FbsA that binds to human Fg . "
ABSTRACT: Interactions of several microbial pathogens with the plasminogen system increase their invasive potential. In this study, we show that Streptococcus agalactiae binds human plasminogen which can be subsequently activated to plasmin, thus generating a proteolytic bacterium. S. agalactiae binds plasminogen via the direct pathway, using plasminogen receptors, and via the indirect pathway through fibrinogen receptors. The glyceraldehyde-3-phosphate dehydrogenase is one of the S. agalactiae proteins that bind plasminogen. Presence of exogenous activators such as uPA and tPA are required to activate bound plasminogen. Results from competitive inhibition assays indicate that binding is partially mediated through the lysine binding sites of plasminogen. Following plasminogen binding and activation, S. agalactiae is able to degrade in vitro fibronectin, one of the host extracellular matrix proteins. Moreover, incubation of S. agalactiae with either plasminogen alone, or plasminogen plus fibrinogen, in the presence of tPA enhanced its virulence in C57BL/6 mice, suggesting that acquisition of plasmin-like activity by the bacteria increase their invasiveness.Microbes and Infection 10/2007; 9(11):1276-84. DOI:10.1016/j.micinf.2007.06.001 · 2.73 Impact Factor
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- "Membrane-bound GAPDH has many functions , but the signal peptide does not have the classical function of catalyzing (Tsai et al. 1982, Allen et al. 1987, Pancholi & Fischetti 1992, Modun & Williams 1999, Argiro et al. 2000a). Surface-located GAPDH of Gram-positive bacteria was reported to be a highly active receptor for host proteins and effective vaccine components (Seifert et al. 2003, Bolton et al. 2004). A 37 kDa surface GAPDH of Schistosoma mansoni is highly antigenic in the immune response of humans against schistosome infection (Argiro et al. 2000a), and further research has demonstrated that GAPDH of S. mansoni carries B-cell epitope and associates with antibody induction (Argiro et al. 2000b) in mice. "
ABSTRACT: Edwardsiella tarda glyceraldehyde-3-phosphate dehydrogenase (GAPDH) may be an effective vaccine candidate against infection by E. tarda in Japanese flounder Paralichthys olivaceus. The GAPDH of E. tarda is highly homologous to that of Vibrio cholerae (91%), and therefore E. tarda GAPDH may have protective antigenicity against Vibrio species. In this study, we immunized Japanese flounder with GAPDH of E. tarda and infected the fish with V anguillarum. The result showed that GAPDH prepared from E. tarda protected Japanese flounder effectively in a challenge of V anguillarum. Therefore, E. tarda GAPDH should be considered as a multi-purpose vaccine candidate against several kinds of pathogenic bacteria.Diseases of Aquatic Organisms 06/2007; 75(3):217-20. DOI:10.3354/dao075217 · 1.59 Impact Factor