A Trigger Enzyme in Mycoplasma pneumoniae: Impact of the Glycerophosphodiesterase GlpQ on Virulence and Gene Expression

Schmidl, Sebastian R.; Otto, Andreas; Lluch‐Senar, María; Piñol, Jaume; Busse, Julia; Becher, Dörte; Stülke, Jörg

doi:10.1371/journal.ppat.1002263

Cited by 65 publications

(80 citation statements)

References 49 publications

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“…The formation of a unique tip-like structure comprising a complex of adhesins and adhesion-related proteins was shown to be the basis for effective adherence of bacteria to the respiratory mucosa (Krause & Balish, 2004). The further colonization process is accompanied by the release of hydrogen peroxide (Hames et al, 2009;Schmidl et al, 2011) and of the pertussis toxin-like CARDS toxin (Kannan & Baseman, 2006), resulting in vacuolization and destruction of epithelial cells.…”

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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Section: Introductionmentioning

confidence: 99%

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

2013

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“…In addition, the number of toxins or other toxic compounds produced by mycoplasmas is limited. Among the known exceptions is the capacity of several mycoplasmas to produce hydrogen peroxide (H 2 O 2 ) as a by-product of glycerol metabolism (13,14). For a small number of Mycoplasma species, the secretion of virulence-mediating proteins was demonstrated.…”

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confidence: 99%

MIB–MIP is a mycoplasma system that captures and cleaves immunoglobulin G

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et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Mycoplasmas are "minimal" bacteria able to infect humans, wildlife, and a large number of economically important livestock species. Mycoplasma infections include a spectrum of clinical manifestations ranging from simple fever to fulminant inflammatory diseases with high mortality rates. These infections are mostly chronic, suggesting that mycoplasmas have developed means to evade the host immune response. Here we present and functionally characterize a two-protein system from Mycoplasma mycoides subspecies capri that is involved in the capture and cleavage of IgG. The first component, Mycoplasma Ig binding protein (MIB), is an 83-kDa protein that is able to tightly bind to the Fv region of a wide range of IgG. The second component, Mycoplasma Ig protease (MIP), is a 97-kDa serine protease that is able to cleave off the VH domain of IgG. We demonstrate that MIB is necessary for the proteolytic activity of MIP. Cleavage of IgG requires a sequential interaction of the different partners of the system: first MIB captures the IgG, and then MIP is recruited to the MIB-IgG complex, enabling protease activity. MIB and MIP are encoded by two genes organized in tandem, with homologs found in the majority of pathogenic mycoplasmas and often in multiple copies. Phylogenetic studies suggest that genes encoding the MIB-MIP system are specific to mycoplasmas and have been disseminated by horizontal gene transfer. These results highlight an original and complex system targeting the host immunoglobulins, playing a potentially key role in the immunity evasion by mycoplasmas.

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“…We aligned the amino acid sequences of GlpQ (amino acids [aa] 298 to 587, excluding the GPDPase_memb domain) and GlpQ2 (aa 1 to 277) with those of GlpQs from H. influenzae and M. pneumoniae, which showed that residues Glu-46, Asp-48, His-61, and Asp-62 in GlpQ2 and Glu-369, Asp-371, His-384, and His-385 in GlpQ were strictly conserved around the active site of GDPD ( Fig. 1) (16,38). This suggests that both GlpQ and GlpQ2 are biologically active enzymes, although only the glpQ gene is found widely in published S. pneumoniae genome sequences.…”

Section: Resultsmentioning

confidence: 99%

“…The cytotoxicity assay was modified from that of Schmidl et al (16). A549 cells were cultured to confluence in a 96-well plate without antibiotics and infected with 5 ϫ 10 6 CFU pneumococci for 2 h at 37°C.…”

Section: Ethics Statementmentioning

confidence: 99%

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Impact of the glpQ2 Gene on Virulence in a Streptococcus pneumoniae Serotype 19A Sequence Type 320 Strain

et al. 2015

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dGlycerophosphodiester phosphodiesterase (GlpQ) metabolizes glycerophosphorylcholine from the lung epithelium to produce free choline, which is transformed into phosphorylcholine and presented on the surfaces of many respiratory pathogens. Two orthologs of glpQ genes are found in Streptococcus pneumoniae: glpQ, with a membrane motif, is widespread in pneumococci, whereas glpQ2, which shares high similarity with glpQ in Haemophilus influenzae and Mycoplasma pneumoniae, is present only in S. pneumoniae serotype 3, 6B, 19A, and 19F strains. Recently, serotype 19A has emerged as an epidemiological etiology associated with invasive pneumococcal diseases. Thus, we investigated the pathophysiological role of glpQ2 in a serotype 19A sequence type 320 (19AST320) strain, which was the prevalent sequence type in 19A associated with severe pneumonia and invasive pneumococcal disease in pediatric patients. Mutations in glpQ2 reduced phosphorylcholine expression and the anchorage of cholinebinding proteins to the pneumococcal surface during the exponential phase, where the mutants exhibited reduced autolysis and lower natural transformation abilities than the parent strain. The deletion of glpQ2 also decreased the adherence and cytotoxicity to human lung epithelial cell lines, whereas these functions were indistinguishable from those of the wild type in complementation strains. In a murine respiratory tract infection model, glpQ2 was important for nasopharynx and lung colonization. Furthermore, infection with a glpQ2 mutant decreased the severity of pneumonia compared with the parent strain, and glpQ2 gene complementation restored the inflammation level. Therefore, glpQ2 enhances surface phosphorylcholine expression in S. pneumoniae 19AST320 during the exponential phase, which contributes to the severity of pneumonia by promoting adherence and host cell cytotoxicity. P hosphorylcholine (ChoP) is a unique feature of many respiratory bacterial species. In the cell wall of pneumococci, ChoP is a component of lipoteichoic acid and teichoic acid that anchors various choline-binding proteins (CBPs) via the choline-binding domain on the pneumococcal surface (1). These CBPs regulate autolysis and the natural transformation of Streptococcus pneumoniae (2), as well as promoting the internalization of pneumococci into pharyngeal epithelial cells to escape phagocytes (3-6).The structure of ChoP resembles that of the platelet-activating factor (PAF), and thus, ChoP on S. pneumoniae can interact directly with the host PAF receptor (7) and allow S. pneumoniae to transit the epithelial and endothelial layers during invasion (8). The conjugation between ChoP and the PAF receptor is important for pneumococcal sequestration during immune clearance and systemic dissemination, since mice deficient in PAF receptor or treated with PAF receptor antagonist abolish pneumococcal pneumonia progression to cause sepsis and meningitis (9).The presentation of ChoP on the bacterial surface requires an available choline source in the environment (10, 11). Fre...

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A Trigger Enzyme in Mycoplasma pneumoniae: Impact of the Glycerophosphodiesterase GlpQ on Virulence and Gene Expression

Cited by 65 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

MIB–MIP is a mycoplasma system that captures and cleaves immunoglobulin G

Impact of the glpQ2 Gene on Virulence in a Streptococcus pneumoniae Serotype 19A Sequence Type 320 Strain

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