Role of secreted glyceraldehyde-3-phosphate dehydrogenase in the infection mechanism of enterohemorrhagic and enteropathogenic Escherichia coli: Interaction of the extracellular enzyme with human plasminogen and fibrinogen

Egea, L.; Aguilera, L.; Gíménez, Rosa; Sorolla, Maria Alba; Aguilar, Juan; Badı́a, Josefa; Baldomà, Laura

doi:10.1016/j.biocel.2007.03.008

Cited by 132 publications

(113 citation statements)

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“…Despite the occurrence of more than 40 cleavage sites for trypsin (www.expasy.ch/ tools/peptidecutter/), proteolytic digestion experiments resulted in a lack of accessible surface-exposed M. pneumoniae GAPDH, whereas M. genitalium GAPDH (82 % identity) was described as trypsin-sensitive (Alvarez et al, 2003). A clear antibody response to recombinant GAPDH was not detected either in hyperimmune sera of (Dallo et al, 2002;Barbosa et al, 2006;Egea et al, 2007), which could influence basic aspects of pathogenesis, like tissue tropism. All three glycolytic enzymes described up to now in M. pneumoniae that act as binding partners of human ECM components are characterized as phosphoproteins , providing an opportunity to regulate the moonlighting role of these proteins via phosphorylation.…”

Section: Discussionmentioning

confidence: 99%

“…Besides their glycolytic activity in the cytosol, these enzymes have been found at the surface of micro-organisms, where they act as binding partners of proteins of the human Abbreviations: ECM, extracellular matrix; FCS, fetal calf serum; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MMR, multiple mutation reaction. Microbiology (2011Microbiology ( ), 157, 2328 As a surface-associated protein, GAPDH can interact with not only one or more specified host factors like fibronectin, fibrinogen, albumin, laminin, collagen and plasminogen but also human colonic mucin, human epithelial and endothelial cells or fimbriae of other bacterial species in biofilms (Pancholi & Fischetti, 1992;Gozalbo et al, 1998;Bergmann et al, 2004;Barbosa et al, 2006;Egea et al, 2007;Kinoshita et al, 2008;Nagata et al, 2009;Purves et al, 2010;Tunio et al, 2010a).Previous studies have confirmed that the described biological function of glycolytic enzymes, in addition to their metabolic role, can also be found in members of the class Mollicutes. Elongation factor Tu and pyruvate dehydrogenase E1 b subunit are located at the surface of M. pneumoniae and were described as binding human fibronectin (Dallo et al, 2002).…”

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Role of Mycoplasma pneumoniae glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in mediating interactions with the human extracellular matrix

2011

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In different, phylogenetically unrelated micro-organisms, glycolytic enzymes play a dual role. In the cytosol they are involved in metabolic reactions whereas the surface-localized fraction of the enzymes contributes to adhesion and virulence. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a typical member of this group of multifunctional proteins. In this study, we characterized the GAPDH of Mycoplasma pneumoniae, a common pathogen of the human respiratory mucosa. Full-length GAPDH of M. pneumoniae was successfully expressed and used to produce a polyclonal antiserum. By immunofluorescence, colony blot and ELISA experiments with different fractions of the M. pneumoniae proteins, GAPDH was demonstrated to be present in the cytosol and at even higher concentrations at the surface of mycoplasmas. Nevertheless, antibodies against recombinant GAPDH were not detected in sera of immunized animals or of patients with confirmed M. pneumoniae infection. Recombinant GAPDH bound to different human cell lines in a concentration-dependent manner, and binding was inhibited by specific anti-GAPDH serum. In contrast, this antiserum did not significantly influence the adherence of M. pneumoniae to HeLa cells. When different human extracellular matrix proteins were tested in Western blot assays, GAPDH bound to fibrinogen. The results showed that the GAPDH of M. pneumoniae is a member of the family of cytosol-localized glycolytic enzymes, which also occur at the surface of the bacterium, and mediates interactions with the extracellular matrix proteins of the human host. Thus, the surface-exposed fraction of GAPDH may be a factor that contributes to the successful colonization of the human respiratory tract by M. pneumoniae. INTRODUCTIONThe cell wall-less bacterium Mycoplasma pneumoniae is an obligate pathogen of the human respiratory tract causing a wide range of different infections. Between 3 and 30 % of all cases of community-acquired pneumonia can be attributed to this micro-organism (Waites & Talkington, 2004). M. pneumoniae mainly infects older children and young adults but recent studies have shown that all age groups can be affected (von Baum et al., 2009). Infections occur endemically in closed populations such as army camps (Klement et al., 2006), but worldwide epidemic peaks at intervals of 3-7 years have been documented (Lind et al., 1997;Eun et al. 2008). Furthermore, respiratory infections can be followed by extra-pulmonary manifestations, causing neurological, gastrointestinal, cardiovascular, haematological and dermatological disorders (Narita, 2010).Despite the reduced genetic repertoire of 814 kb and the limited metabolic pathways (Himmelreich et al., 1996;Dandekar et al., 2000), M. pneumoniae is perfectly adapted to interaction with the human respiratory mucosa, which is the only known infection site. The unique tip structure (attachment organelle) of these mycoplasmas comprises a complex network of adhesins and adherence-associated proteins that mediate the directed adhesion of the bacteria to the epitheli...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Role of Mycoplasma pneumoniae glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in mediating interactions with the human extracellular matrix

2011

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“…Isolation of secreted proteins was performed as previously described (Egea et al, 2007). Briefly, overnight cultures in LB were diluted 1:50 in the indicated culture media and incubated without shaking at 37ºC in a 5% CO 2 atmosphere.…”

Section: Protein Secretion Assaysmentioning

confidence: 99%

“…In these pathogens, secretion is not linked to outer membrane vesicles and depends on growth conditions. The protein is secreted at 37°C by cells grown in LB or in eukaryotic culture media such as DMEM or Ham's F-12, but not in glucose-minimal medium (Egea et al, 2007). E. coli GAPDH binds human plasminogen and fibrinogen and remains associated with Caco-2 cells upon infection (Egea et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Secretion of the housekeeping protein glyceraldehyde-3-phosphate dehydrogenase by the LEE-encoded type III secretion system in enteropathogenic Escherichia coli

Aguilera

Ferreira

Gíménez

et al. 2012

The International Journal of Biochemistry & Cell Biology

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“…The dual role of glycolytic enzymes was first described in streptococci (Pancholi & Fischetti, 1992). Meanwhile, these interactions have been reported for phylogenetically different micro-organisms, including fungi, parasites, and Gram-positive and Gramnegative bacteria (Barbosa et al, 2006;Bergmann et al, 2003;Egea et al, 2007;Gozalbo et al 1998;Lama et al, 2009). Furthermore, an increasing number of microbial glycolytic enzymes have been characterized as involved in the interaction with human ECM, such as glyceraldehyde-3-phosphate dehydrogenases (Pancholi & Fischetti, 1992) and enolases (Pancholi & Fischetti, 1998).…”

Section: Introductionmentioning

confidence: 99%

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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Role of secreted glyceraldehyde-3-phosphate dehydrogenase in the infection mechanism of enterohemorrhagic and enteropathogenic Escherichia coli: Interaction of the extracellular enzyme with human plasminogen and fibrinogen

Cited by 132 publications

References 36 publications

Role of Mycoplasma pneumoniae glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in mediating interactions with the human extracellular matrix

Role of Mycoplasma pneumoniae glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in mediating interactions with the human extracellular matrix

Secretion of the housekeeping protein glyceraldehyde-3-phosphate dehydrogenase by the LEE-encoded type III secretion system in enteropathogenic Escherichia coli

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

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