Role of the C-Terminal Lysine Residues of Streptococcal Surface Enolase in Glu- and Lys-Plasminogen-Binding Activities of Group A Streptococci

Derbise, Anne; Song, Youngmia P.; Parikh, Sonia; Fischetti, Vincent A.; Pancholi, Vijay

doi:10.1128/iai.72.1.94-105.2004

Cited by 106 publications

(135 citation statements)

References 49 publications

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“…The binding region (amino acids 91-102) is located directly after the predicted transmembrane domain of the protein (www.cbs.dtu.dk/services/TMHMM/). M. pneumoniae PDHB shows a C-terminal lysine which was characterized as a potential binding region in enolase (Derbise et al, 2004), but no interaction of the corresponding recombinant protein derived from this region with human plasminogen was found. In contrast, the plasminogen-binding peptide identified has no lysine, indicating lysineindependent interactions between the two proteins.…”

Section: Discussionmentioning

confidence: 99%

“…In most cases, enolases interact with plasminogen but binding to fibronectin and laminin has also been reported (Carneiro et al, 2004;Castaldo et al, 2009;Donofrio et al, 2009). The M. pneumoniae enolase shows typical putative plasminogen-binding sites, such as lysine in the C terminus, 448 FKNIK 452 , and a lysine-rich internal motif, 268 KRYVFKKGIKAKILDEK 284 (Bergmann et al, 2003;Derbise et al, 2004;Yavlovich et al, 2007). A surprising fact was the absence of this enzyme on the surface of mycoplasma cells.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of pyruvate dehydrogenase subunit B and enolase as plasminogen-binding proteins in Mycoplasma pneumoniae

2013

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The obligate pathogenic mycoplasma species Mycoplasma pneumoniae uses a limited but effective repertoire of virulence factors to infect and colonize the human respiratory tract. Besides the development of a unique adhesion complex and the expression of tissue-damaging factors, surface-located glycolytic enzymes and their capacity to bind to components of the human extracellular matrix (ECM) support pathogen-host interactions. Here, we demonstrated that the glycolytic enzymes enolase (Mpn606) and pyruvate dehydrogenase subunit B (Mpn392; PDHB) of M. pneumoniae show concentration-dependent binding to human plasminogen. Monospecific polyclonal antisera against both recombinant proteins reduced the binding to plasminogen significantly. The surface location of PDHB but not of enolase was demonstrated using Triton X fractionation of M. pneumoniae total protein content, membrane fractionation, colony blotting, mild proteolysis of mycoplasma cells, and immunofluorescence tests. To characterize the binding site of plasminogen in surface-displaced PDHB, the mycoplasmal protein was separated into four recombinant proteins followed by investigation of the binding behaviour of peptides that overlap the protein part interacting with plasminogen. Spot analysis resulted in a novel region of 12 amino acids (FPAMFQIFTHAA, position 91 to 102 of PDHB), which is responsible exclusively for binding of human plasminogen and also interacts in a dose-dependent manner with this host protein. The data indicate that the plasminogen-binding enzymes enolase and especially the surface-associated PDHB may contribute to the pathogenesis of M. pneumoniae infections.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Characterization of pyruvate dehydrogenase subunit B and enolase as plasminogen-binding proteins in Mycoplasma pneumoniae

2013

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“…In addition, PspC and the PspClike protein Hic have been shown to interact with complement factor H (Dave et al, 2004;Janulczyk et al, 2000). Many pathogenic bacteria also produce receptors for Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GMFI, geometric mean fluorescence intensity; immuno-FESEM, field emission scanning immunoelectron microscopy; PspA, PspC, pneumococcal surface protein A or C. The plasminogen-binding motifs in the enolase enzyme were initially traced to C-terminal lysine residues (Bergmann et al, 2001(Bergmann et al, , 2004aDerbise et al, 2004). However, a subsequent study identified a nine residue internal plasminogen-binding site in enolase .…”

Section: Introductionmentioning

confidence: 99%

Streptococcus pneumoniae enolase is important for plasminogen binding despite low abundance of enolase protein on the bacterial cell surface

et al. 2006

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Enolase represents one of the anchorless surface proteins of Streptococcus pneumoniae and has previously been identified as a plasminogen-binding protein, endowing this pathogen with host proteolytic activity. In this study the mAb 245,C-6 (IgG1) was produced in a BALB/c mouse after immunizing with a protein fraction from S. pneumoniae. The mAb reacted with recombinant pneumococcal enolase both under non-denaturing and denaturing conditions. The epitope for the mAb was mapped to residues 55 DKSRYGGLG 63 of pneumococcal enolase using a peptide array. By applying the previously reported structure of enolase, this epitope was localized in a surface-exposed loop in each of the monomers of the octameric enolase. Previous immunoelectron microscopic studies, using polyclonal rabbit antibodies against enolase, depicted enolase on the cell surface but did not quantify the amount of surface-exposed enolase on viable pneumococci. Here, flow cytometry revealed no binding of mAb 245,C-6 to viable pneumococci, including TIGR4 and its non-encapsulated isogenic mutant, and only a minor increase of fluorescence intensity was measured when the polyclonal anti-enolase antibodies were used. In contrast, control antibodies recognizing the choline-binding proteins (CBPs) PspA and PspC showed high reactivities. The non-encapsulated TIGR4 did not show increased levels of antibody binding for mAb 245,C-6 or polyclonal anti-enolase antibodies, but revealed increased binding of polyclonal antibodies reacting with PspA or PspC. These results suggest that, compared to other surface-exposed proteins such as CBPs, the amount of enolase under the selected conditions is low. Flow cytometry, however, with FITC-labelled plasminogen demonstrated that the amount of surface-exposed enolase is important for plasminogen binding and, therefore, is also important for pneumococcal pathogenesis. INTRODUCTIONStreptococcus pneumoniae is an important human pathogen causing a variety of diseases including pneumonia, meningitis, sepsis and otitis media. A prerequisite for invasive pneumococcal diseases (infections) is the ability of the bacteria to colonize the mucosal surface and penetrate the mucosal barriers. During these processes binding of bacterial factors such as adhesins are essential for the interactions with receptors on host cells. Among the bacterial factors that mediate direct adherence to human cells are phosphorylcholine (Cundell et al., 1995) and the choline-binding protein PspC, also known as SpsA and CbpA (Brooks-Walter et al., 1999;Hammerschmidt et al., 1997;Rosenow et al., 1997). The PspC protein binds to the secretory component of the polymeric Ig receptor or secretory IgA and this interaction mediates transmigration of pneumococci through mucosal epithelial cells (Elm et al., 2004;Zhang et al., 2000). In addition, PspC and the PspClike protein Hic have been shown to interact with complement factor H (Dave et al., 2004;Janulczyk et al., 2000). Many pathogenic bacteria also produce receptors for Abbreviations: GAPDH, glyceraldehyde-3-phos...

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“…Remarkably, the analyses have identified a significant number of traditional cytoplasmic proteins commonly thought to be intracellular in streptococci. Many of these proteins have been identified as virulence factors in streptococci, such as the glycolytic pathway enzymes enolase and GAPDH, which have been identified as plasminogen binding and cell wall-associated in S. pyogenes [125][126][127] and in S. pneumoniae [128].…”

Section: Anchorless Proteinsmentioning

confidence: 99%

Human pathogenic streptococcal proteomics and vaccine development

Cole

Henningham

Gillen

et al. 2008

Proteomics Clinical Apps

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Gram-positive streptococci are non-motile, chain-forming bacteria commonly found in the normal oral and bowel flora of warm-blooded animals. Over the past decade, a proteomic approach combining 2-DE and MS has been used to systematically map the cellular, surface-associated and secreted proteins of human pathogenic streptococcal species. The public availability of complete streptococcal genomic sequences and the amalgamation of proteomic, genomic and bioinformatic technologies have recently facilitated the identification of novel streptococcal vaccine candidate antigens and therapeutic agents. The objective of this review is to examine the constituents of the streptococcal cell wall and secreted proteome, the mechanisms of transport of surface and secreted proteins, and describe the current methodologies employed for the identification of novel surface-displayed proteins and potential vaccine antigens.

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Role of the C-Terminal Lysine Residues of Streptococcal Surface Enolase in Glu- and Lys-Plasminogen-Binding Activities of Group A Streptococci

Cited by 106 publications

References 49 publications

Characterization of pyruvate dehydrogenase subunit B and enolase as plasminogen-binding proteins in Mycoplasma pneumoniae

Characterization of pyruvate dehydrogenase subunit B and enolase as plasminogen-binding proteins in Mycoplasma pneumoniae

Streptococcus pneumoniae enolase is important for plasminogen binding despite low abundance of enolase protein on the bacterial cell surface

Human pathogenic streptococcal proteomics and vaccine development

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